~【NEW!!活動快報】~ 全景玻片影像掃描器,精準對焦與掃描,染色結果從此數位化
MoticEasyScan 全景玻片影像掃描器,3D多層次掃描,快速對焦精準掃描為數位玻片。提供最佳解析度0.25um/pixel ;6片式片夾,walk-away設計。搭配簡易操作軟體,優質影像數位永久保存。歡迎來電洽詢(02)86609496許虹宜

目前分類:【細胞轉染、培養、螢光封片劑】 (26)

瀏覽方式: 標題列表 簡短摘要

DAPI螢光染劑4',6-二脒基-2-苯基吲哚

 

品牌ABMGOOD/貨號B551/ 包裝0.2ML(0.5mg/mL)

DAPI 藍色螢光螢劑,是一種能與DNA強力結合的螢光染劑當染劑結合至dsDNA AT regions時,螢光會呈現20倍強度。最大吸收波長為364nm/散射波長454nm (藍色)DAPI常用於顯微鏡觀察,螢光染劑可以透過完整的細胞膜,可用於活細胞 (高濃度時) 和固細胞的染色。

 

DAPI (4',6-diamidino-2-phenylindole) is used extensively as a nuclear stain that binds strongly to A-T rich regions in the minor groove of double-stranded DNA. The blue fluorescence increases 20-fold when DAPI binds to dsDNA. Since it can pass through cell membrane, DAPI can be used to stain both live and fixed cells, but much higher concentration has to be used for live cells as its permeability for live cell membrane is dramatically less.

DAPI4',6-二脒基-2-苯基吲哚(4',6-diamidino-2-phenylindole)

DAPI產品應用

1.     黴漿菌污染偵測

2.     免疫螢光染色

3.     流氏細胞儀

4.     細胞核染劑

 

DAPI產品提供 (產品貨號#B551)

0.2ml DNA染劑 (5mg/ml)

 

DAPI 內容相關

內容相關https://www.abmgood.com/DAPI-(4,6-Diamidino-2-Phenylindole,-Dihydrochloride)-B551.html#1

 

 

HOT!! 其它黴漿菌相關商品如下連結

 

檢測/黴漿菌PCR偵測商品

檢測黴漿菌DAPI偵測商品

◆移除/細胞內黴漿菌除去商品 (細胞內)

預防培養箱添加-黴漿菌除去商品 (培養箱設備)

預防水浴槽添加-黴漿菌除去商品 (水浴槽設備)

預防噴霧式黴漿菌除去商品 (細胞培養設備)

 

 

 

更多相關問題洽詢

------------------------------

台灣代理廠商: 太鼎生物科技有限公司

公司電話: 02-86609496 /0920312382
公司傳真: 02-86609342
公司網站: www.biopioneer.com.tw
連絡業務: 許虹宜 0920312382
連絡信箱: taiding.biotech@msa.hinet.net

文章標籤

太鼎生物科技 發表在 痞客邦 留言(0) 人氣()

DNAfectin™ 2100 Transfection Reagent

強效型DNA轉染試劑

 


DNAfectin™ Transfection Reagents

1. 強效型DNA轉染試劑。

2. 獨特陽離子結合liposomes,大幅提高DNA於真核細胞轉染效果。

 

3. 細胞繼代15代後,仍有高效轉染效果。

 

4. 針對難轉的懸浮細胞、初代細胞、神經細胞都有高效表現。

5. 提供完整各類細胞轉染方法。

 

 

 



















Cat.No.:




G2100




Quantity:




1.0mg/1.0ml




 




 





Data
Sheet


































































































































Description




DNAfectin™ 2100 Transfection Reagent
is a unique formulation of multiple polycations and liposomes that enable
highly efficient and effective DNA transfection of eukaryotic cells.
DNAfectin™ is the best transfection reagent for over 15 different cell
lines
, including difficult-to-transfect, suspension, and primary cells.
This transfection reagent is perfect for any of your transfection needs.




Application




Transfection of DNA into cultured
eukaryotic cells.




Protocol



























Kit Contents




1ml at 1.0mg/1.0ml




Storage




Store at 4ºC. Do not freeze.




Notes




Please check our Transfection Cell
Database for tested cell lines




Cell
Line




Cell Line Species/Tissue




Nucleic Acid




Reagent




Transfection Type




Transfection Efficiency(%)




293T




Human Embryonic Kidney




DNA




DNAfectinTM
2100 Transfection Reagent




Transient




100%




Jurkat




Human T Lymphoma




DNA




DNAfectinTM
2100 Transfection Reagent




Transient




25%




SHEP




Human Neuroblastoma




DNA




DNAfectinTM
2100 Transfection Reagent




Transient




90%




BOSC23




Human Kidney




DNA




DNAfectinTM
2100 Transfection Reagent




Transient




90%




HT1080




Human Fibrosarcoma




DNA




DNAfectinTM
2100 Transfection Reagent




Transient




80%




C33A




Human Cervical Carcinoma




DNA




DNAfectinTM
2100 Transfection Reagent




Transient




90%




MRC5




Human Primary Fibroblast




DNA




DNAfectinTM
2100 Transfection Reagent




Transient




80%




MDK-Telo




Dog Kidney Cells+Telo




DNA




DNAfectinTM
2100 Transfection Reagent




Transient




80%




B16




Murine Melanoma




DNA




DNAfectinTM
2100 Transfection Reagent




Transient




80%




CB3




Mouse Erytholeukemia




DNA




DNAfectinTM
2100 Transfection Reagent




Transient




25%




HuVEC




Human Ubilical Vein




DNA




DNAfectinTM
2100 Transfection Reagent




Transient




20%




HepG2




Human Liver Carcinoma




DNA




<p style="margi

太鼎生物科技 發表在 痞客邦 留言(0) 人氣()


Cell Line Lysates


 


























































































































































Product




Description




Cat No.




Price



























Neuro 2A




Mouse Neuroblastoma; 100ug/100ul




L006




$65.00







NIE115




Mouse
Neuroblastoma; 100ug/100ul




L007




$65.00







SH-Sy5y




Human
Neuroblastoma; 100ug/100ul




L008




$65.00







U87MG




Human
Glioblastoma; 100ug/100ul




L009




$65.00







TE671




Human
Medulloblastoma; 100ug/100ul




L010




$65.00







SHEP




Human
Neuroblastoma; 100ug/100ul




L011




$65.00







ABMC1




Human
Embryonic Retinoblast; 100ug/100ul>




L012




$65.00







IMR-90




Human
Fetus Lung Fibroblast; 100ug/100ul




L013




$65.00







MRC-5




Human
Fetus Lung Fibroblast; 100ug/100ul




L014




$65.00







WI-38




Human Fetus Lung Fibroblast;
100ug/100ul




L015




$65.00







LLC1




Mouse Lewis Lung Carcinoma;
100ug/100ul




L016




$65.00







H441




Human
Lung Papillary Adenocarcinoma; 100ug/100ul




L017




$65.00







Calu-6




Human
Lung Carcinoma; 100ug/100ul




L018




$65.00







MDA-MB-468




Human
Breast Adenocarcinoma; 100ug/100ul




L019




$65.00







MDA-MB-231




Human
Breast Adenocarcinoma; 100ug/100ul




L020




$65.00







MCF7




Human
Breast Adenocarcinoma; 100ug/100ul




L021




$65.00







SW-48




Human
Colorectal Adenocarcinoma; 100ug/100ul




L022




$65.00







T-47D




Human
Breast Ductal Carcinoma; 100ug/100ul




L023




$65.00







Hep3B




Human
Hepatocellular Carcinoma; 100ug/100ul




L024




$65.00







HepG2




Human
Hepatocellular Carcinoma; 100ug/100ul




L025




$65.00







HCT15




Human
Colorectal Adenocarcinoma; 100ug/100ul




L026



太鼎生物科技 發表在 痞客邦 留言(0) 人氣()

 



Cell Lysates & Controls




Tag Lysates




表單的





































































Product




Description




Cat No.




Price




GST
Lysates




293 cells transfected with GST minigene, 26 kDa.
100µg/100µl




L004




$65.00




Myc
Lysates




293 cells transfected with Myc-tagged minigene,
37 kDa. 100µg/100µl




L001




$65.00




His
Lysates




293 cells transfected with His-tagged minigene,
50 kDa. 100µg/100µl




L003




$65.00




HA Lysates




293 cells transfected with HA-tagged minigene, 55
kDa. 100µg/100µl




L002




$65.00




D-Tag
Lysates




293 cells transfected with D-Tag-tagged minigene,
60 kDa. 100µg/100µl




L074




$65.00




GFP
Lysates




293 cells transfected with GFP minigene, 30 kDa.
100µg/100µl




L079




$65.00




V5 Lysates




293 cells transfected with V5-tagged fusion
constructs, 130 kDa. 100µg/100µl




L082




$65.00




β-Gal
Lysates




293 cells transfected with β-Gal-tagged fusion
constructs, 130 kDa. 100µg/100µl




L081




$65.00




VSVG
Lysates




293 cells transfected with a VSVG construct, 50
kDa. 100µg/100µl




L080




$65.00





太鼎生物科技 發表在 痞客邦 留言(0) 人氣()




Amphotericin B (Fungizone)




Antibiotic
Solutions




Amphotericin B (Fungizone) (100X)




Amphotericin
B (Fungizone)
































Cat.No.:




G274




Quantity:




100ml




Price:




請電洽02-86609496




Description




This product contains 250 µg/ml of
amphotericin B and 205 µg/ml of deoxycholic acid in normal saline. It is 0.2
micron filtered and is tested for sterility.




Storage




This product is stored at -20°C . At 37°C , it is stable for up to 7 days. At 2 -8°C , it is stable for up to 3 weeks. Avoid
freeze-thaw cycles.




Data Sheet


























Product Description




This product contains 250 μg/ml of amphotericin B and
205
μg/ml of deoxycholic acid in normal saline. It is
0.2 micron filtered and is tested for sterility.




Usage




Amphotericin B is an antifungal agent used in cell cultures. It should be
added to cell cultures at a maximum concentration of 10ml per litre of medium.




Precautions and Disclaimer




For laboratory use only. Not for drug, household or other uses.




Storage




This product is stored at -20oC. At 37oC, it is stable for up to 7 days. At 2-8oC, it is stable for up to 3 weeks. Avoid freeze-thaw cycles.




太鼎生物科技 發表在 痞客邦 留言(0) 人氣()





Geneticin (G418)





抗生素G418, Geneticin (Antibiotic G418)





 Geneticin Solution (Antibiotic G418) (50mg/ml)

































Cat.No.:




G271




Quantity:




10ml




Price:




請電洽02-86609496




Description




This
product contains 50 mg/ml of active G418. It is 0.2 micron filtered and is
tested for sterility.




Storage




This
product is stored at 2 -8°C
and is stable for 2 years.




Data Sheet



























Product Description





This product contains 50 mg/ml of active G418. It is 0.2 micron filtered and
is tested for sterility.





Usage





The aminoglycoside antibiotic G418 is effective against both eukaryotic
and prokaryotic cells. It is used to select for cells that express neomycin
resistance as a result of successful transfection. G418 should be added to
animal cell cultures at a concentration of 400
μg/ml for selection
purposes, while a concentration of 200
μg/ml is
sufficient for maintenance. For prokaryotic cells, the maximum con­centration
is 5
μg/ml.





Precautions and Disclaimer





For laboratory use only. Not for drug, household or other uses.





Storage





This product is stored at 2-8oC and is stable for 2 years.




太鼎生物科技 發表在 痞客邦 留言(0) 人氣()




青黴素/鏈黴素雙抗生素溶液, Penicillin/Streptomycin Solution




Penicillin/Streptomycin




 Penicillin-Streptomycin Solution (100X)
































Cat.No.:




G255




Quantity:




100ml




Price:




請電洽02-86609496




Description




Penicillin-
Streptomycin contains 10,000 units of penicillin (base), 10,000µg of
streptomycin (base) per mL in WFI water. It is supplied as a 0.22
µm-filtered, 100X frozen liquid.




Storage




This
product is stable for 2 years at -20°C .




Data Sheet


























 




 Product Description




Penicillin- Streptomycin contains10,000 units of penicillin (base), 10,000
μg of
streptomycin (base) per mL in WFI water. It is supplied as a 0.22
μm-filtered, 100X frozen liquid.




Usage




As a 100X concentration, it is designed to be supplemented as 1 ml per 99
ml of basal medium, yielding an iso-osmotic solution at final dilution.




Storage




This product is stable for 2 years at -20oC.




太鼎生物科技 發表在 痞客邦 留言(0) 人氣()





Antibiotic Solutions





抗生素溶液Gentamicin





Gentamicin Solution (50mg/ml), 10ml





Gentamicin

































Cat.No.:




G272




Quantity:




10ml




Price:




請電洽02-86609496




Description




This
product contains 50 mg/ml of active gentamicin sulfate in WFI water. It is
0.2 micron filtered and is tested for sterility.




Storage




This
product is stored at 2 -8°C .
At room temperature, this product should be consumed within two days. It is
stable at 37oC for five days.




Data Sheet



























Product Description





This product contains 50 mg/ml of active gentamicin sulfate in WFI water.
It is 0.2 micron filtered and is tested for sterility.





Usage





Gentamicin has the advantage of being a broad-spectrum antibiotic that is
non-toxic to animal cells and viruses at recommended concentrations. It should
be added to cell cultures at a maximum concentration of 1ml per litre, with a
working concentration of 50
μg/mL for eukaryotic cell culture and 15 μg/mL for prokaryotic cell culture.





Precautions and Disclaimer





For laboratory use only. Not for drug, household or other uses.





Storage





This product is stored at 2-8oC. At room temperature, this product should be consumed within two days.
It is stable at 37
oC for five days.




太鼎生物科技 發表在 痞客邦 留言(0) 人氣()




 


胎牛血清 Cell Culture Products




Research
Grade Origin




100ml | 500ml




Dialyzed




100ml | 500ml




Charcoal
Stripped




100ml | 500ml




 




USDA Research Grade Origin Fetal
Bovine Serum




 




































Cat.No.:




TM999-500




Quantity:




500ml




Price:




請電洽02-86609496




Description




The USDA
research grade origin fetal bovine serum is an economical alternative to more
stringently purified serums. It contains a maximum endotoxin level of 20EU/ml
and a maximum hemoglobin level of 30mg/dl. This product is best suited for
routine cell culture work.




Sterility




This USP
sterility tested product has been triple 0.1µm filtered. It has been screened
for mycoplasma and viruses, and is certified to be free of bovine diarrhea
virus, cytopathogenic agents, and hemoadsorbing agents.




Storage




This
product is stored at -20°C ,
with a shelf life is 60 months. Repeated freeze-thaw cycles should be avoided
by making aliquots upon first thawing. Like most serum products, turbidity
may occur upon thawing. This does not affect the performance of this product.




Data Sheet


























USDA Dialyzed Fetal Bovine Serum




































Cat.No.:




TM997-500




Quantity:




500ml




Price:




請電洽02-86609496




Description




The USDA
dialyzed fetal bovine serum is a highly purified serum designed for the
investigation of alternative biochemical pathways. The levels of small
molecules such as amino acids and nucleosides have been minimized using a
dia-filtration process. This product contains a maximum endotoxin level of
10EU/ml and a maximum hemoglobin level of 20mg/dl.




Sterility




This USP
sterility tested product has been dialyzed using a 10,000MW cutoff filter
against normal saline. The concentrations of hypoxanthine and thymidine have
been reduced below detectable levels. The level of glucose, which is
indicative of the extent of small molecule removal, has a maximum of 5mg/dl.
This product has also been screened for mycoplasma and viruses, and it is
certified to be free of bluetongue virus, bovine adenovirus, bovine
parvovirus, bovine respiratory syncytial virus, bovine diarrhea virus, rabies
virus, reovirus, cytopathogenic agents, and hemoadsorbing agents.




Storage




This
product is stored at -20°C ,
with a shelf life is 60 months. Repeated freeze-thaw cycles should be avoided
by making aliquots upon first thawing. Like most serum products, turbidity
may occur upon thawing. This does not affect the performance of this product.




Data Sheet








USDA Charcoal Stripped Fetal Bovine Serum




































Cat.No.:




TM998-500




Quantity:




500ml




Price:




請電洽02-86609496




The USDA
charcoal stripped fetal bovine serum is a highly purified serum designed for
specialized cell culture work. The levels of hormones, growth factors, and
steroids have been minimized, making this product ideal for the study of cell
signalling. This product contains a maximum endotoxin level of 10EU/ml and a
maximum hemoglobin level of 20mg/dl.



 



Sterility




This USP
sterility tested product has been triple 0.1µm filtered. It has been screened
for mycoplasma and viruses, and is certified to be free of bluetongue virus,
bovine adenovirus, bovine parvovirus, bovine respiratory syncytial virus,
bovine diarrhea virus, rabies virus, reovirus, cytopathogenic agents, and
hemoadsorbing agents.




Storage




This product
is stored at -20°C ,
with a shelf life is 60 months. Repeated freeze-thaw cycles should be avoided
by making aliquots upon first thawing. Like most serum products, turbidity
may occur upon thawing. This does not affect the performance of this product.




Data Sheet








太鼎生物科技 發表在 痞客邦 留言(0) 人氣()




 


Prigrow Medium




Type I | Type II | Type III |
Type IV




Prigrow I Medium


























Cat.No.:




TM001




Quantity:




500ml




Price:




請電洽02-86609496




Description




Sterile-filtered,
endotoxin free and cell culture tested medium for primary cells.




Storage




Store at 2 -8°C .





Prigrow II Medium


























Cat.No.:




TM002




Quantity:




500ml




Price:




請電洽02-86609496




Description




Sterile-filtered,
endotoxin free and cell culture tested medium for primary cells.




Storage




Store at 2 -8°C .





Prigrow III Medium


























































Cat.No.:




TM003




Quantity:




500ml




Price:




請電洽02-86609496




Description




Sterile-filtered,
endotoxin free and cell culture tested medium for primary cells.




Appearance(Turbidity)




Clear




Appearance(Form)




Solution




pH




7.0 - 7.6




Osmolality




327 - 361
mOs/kg




Cell
Culture Testing-MTT




Pass




Endotoxin
Level




<= 1
EU/ml




Glucose
Concentration




4.05 - 4.95 g /l




Sterility




Pass




Storage




Store at 2 -8°C





Prigrow IV Medium



























Cat.No.:




TM004




Quantity:




500ml




Price:




請電洽02-86609496




Sterile-filtered,
endotoxin free and cell culture tested medium for primary cells.



 



Storage




Store at 2 -8°C .



 




太鼎生物科技 發表在 痞客邦 留言(0) 人氣()




 


Applied
Cell Extracellular Matrix




Applied Cell
Extracellular Matrix


















Cat.No.:




G422




Quantity:




25ml




Price:




請電洽02-86609496





Description




In cell biology, processes such as cell
differentiation, cell cycle and cell survival are affected by signals
stimulated through cell adhesion. Therefore adhesion plays a fundamental role
in in vitro studies. ABM's Applied Cell Extracellular Matrix composes mainly of
type I collagen, isolated from rat tail tendon. Type I collagen is the most
abundant form of collagen and differs from other collagens by its low lysine
hydroxylation and low carbohydrate composition. This product is suitable for
usage as a solid gel or for coating of culture wares. ABM's Applied Cell
Extracellular matrix on culture surfaces have been shown to enhance cell
attachment in a wide variety of cell lines such as hepatocytes, fibroblasts and
epithelial cells.




Features




-sterile liquid containing 4mg/ml of pure
collagen (>95%).

-stored in 20 mM
acetic acid (pH ~3.0).

-direct addition to culture wares for coating

-simple and rapid plate coating procedure.

-enhance cell attachment in a variety of primary cells.




Storage




4-8ºC




Data Sheet






















太鼎生物科技 發表在 痞客邦 留言(0) 人氣()




 


Applied
Cell Extracellular Matrix




Applied Cell
Extracellular Matrix


















Cat.No.:




G422




Quantity:




25ml




Price:




請電洽02-86609496





Description




In cell biology, processes such as cell
differentiation, cell cycle and cell survival are affected by signals
stimulated through cell adhesion. Therefore adhesion plays a fundamental role
in in vitro studies. ABM's Applied Cell Extracellular Matrix composes mainly of
type I collagen, isolated from rat tail tendon. Type I collagen is the most
abundant form of collagen and differs from other collagens by its low lysine
hydroxylation and low carbohydrate composition. This product is suitable for
usage as a solid gel or for coating of culture wares. ABM's Applied Cell
Extracellular matrix on culture surfaces have been shown to enhance cell
attachment in a wide variety of cell lines such as hepatocytes, fibroblasts and
epithelial cells.




Features




-sterile liquid containing 4mg/ml of pure
collagen (>95%).

-stored in 20 mM
acetic acid (pH ~3.0).

-direct addition to culture wares for coating

-simple and rapid plate coating procedure.

-enhance cell attachment in a variety of primary cells.




Storage




4-8ºC




Data Sheet






















太鼎生物科技 發表在 痞客邦 留言(0) 人氣()



L-glutamine 200mM Solution (100X)


 



 




100倍200mM麩醯胺酸溶液


 


L-glutamine




Product Description




This product contains 29.2
g /L of L-glutamine in normal saline. It is 0.2 micron filtered
and is tested for sterility. While L-glutamine is most stable at 4
oC and neu­tral pH, it can spontaneously degrade even under these
conditions. L-glutamine should be kept at low temperatures to minimize the
formation of ammonia as a degradation product.




Uses




The essential amino acid L-glutamine is used in energy production, protein
syn­thesis, and nucleic acid generation by most animal and insect cells. This
product can be used as a nutrient supplement in cell cultures.




Precautions and Disclaimer




For laboratory use only. Not for drug, household or other uses.




Storage




This product is stored at -20oC, with a shelf life of two years. Once thawed, the product should be used
within two weeks.
































Cat.No.:




G275




Quantity:




100ml




Price:




請電洽02-86609496




Description




This product
contains 29.2 g /L
of L-glutamine in normal saline. It is 0.2 micron filtered and is tested for
sterility. While L-glutamine is most stable at 4oC and neutral pH, it can
spontaneously degrade even under these conditions. L-glutamine should be kept
at low temperatures to minimize the formation of ammonia as a degradation
product.




Storage




This
product is stored at -20°C ,
with a shelf life of two years. Once thawed, the product should be used
within two weeks.




Data Sheet


























 




太鼎生物科技 發表在 痞客邦 留言(0) 人氣()



Cell Culture Products


 





 


胰蛋白酶 (細胞培養)


Trypsin-EDTA 0.25%
Solution




100ml | 500ml




Trypsin-EDTA
































Cat.No.:




TM050




Quantity:




500ml




Price:




電浛02-86609496 許虹宜




Description




Trypsin-EDTA
(0.25%) contains 2.5 g /L
Porcine Trypsin (tested for porcine parvovirus) and 0.2g /L EDTA-Na in Hanks Balanced Salt
Solution, with Phenol Red and without Ca
2+ and Mg 2+.




Storage




This
product is stable for 2 years at -20°C .




Data Sheet



























 


Trypsin-EDTA 0.25%
Solution




Store at -20°C




Product Description




Trypsin-EDTA (0.25%)
contains 2.5 g /L Porcine Trypsin
(tested for porcine parvovi­rus) and 0.2g /L
EDTA
4Na in Hanks’ Balanced Salt Solution, with Phenol Red and without
Ca
2+ and Mg 2+.




Usage




Trypsin-EDTA (0.25%)
is recommended for detachment of adherent cells, enzy­matic dissociation of
tissue into single cell suspensions, dissociation of ES cell colonies, and
dissociation of embryoid bodies.




Precautions and Disclaimer




For laboratory use
only. Not for drug, household or other uses.




Storage




This product is
stable for 2 years at -20
oC.




太鼎生物科技 發表在 痞客邦 留言(0) 人氣()

 


             細胞分析產品介紹


一、  免疫螢光染色(Immunofluorescence, IF)


1.      Lightning-link (directly labeling kit) 抗體直接螢光標誌。


具有各種顏色抗體標誌套組。當多重抗體染色時,不須考慮host來源的問題、避免二抗所造成非專一性結合。


 


更多參考資料如下所示:


分類:Lighting-link抗體、蛋白質螢光標誌


http://tw.myblog.yahoo.com/taiding2000/article?mid=4&prev=5&l=f&fid=38


 


2. 螢光封片劑 (mounting solution)


專為免疫螢光實驗所設計,內含螢光保護劑,可大幅減緩螢光分子氧化褪色的情形。


 


更多參考資料如下所示:


分類:螢光封片劑、呈色試劑、EIA分析套組


http://tw.myblog.yahoo.com/taiding2000/article?mid=38&prev=45&l=f&fid=18


 


3. DAPI螢光封片劑 (mounting solution with DAPI dye)


DAPI螢光封片劑, 可以染核及封片同時進行, 不管您先前DAPI是後染5分鐘或是與二抗一起共染,現在都不用。 在最後封片時, 就完成染核動作!!


專為免疫螢光實驗所設計,內含螢光保護劑,可大幅減緩螢光分子氧化褪色的情形。


 


更多參考資料如下所示:


分類:螢光封片劑、呈色試劑、EIA分析套組


http://tw.myblog.yahoo.com/taiding2000/article?mid=38&prev=45&l=f&fid=18


 


二、  細胞轉染與感染 (cell transfection)


 


1. T-Pro Plasmid Midi Kits


T-Pro midi 質體DNA萃取套組 (plasmid midi extraction kit)


是一種利用陰離子交換樹脂純化中量菌液(250毫升)中質體或載體DNA之實驗套組。利用此一套組所純化之DNA流程可於2小時內完成並無須其他額外處理即可進行。純化出來的質體DNA,適用於後續實驗進行,例如;細胞轉染、限制酶之處理、DNA接合、聚合酶連鎖反應、試管內轉錄及定序反應等。


 


更多參考資料如下所示:


分類:DNA、PCR、QPCR分生試劑http://tw.myblog.yahoo.com/taiding2000/article?mid=38&prev=45&l=f&fid=18


 


2. NTRII細胞轉染試劑 (NTRII cell transfection reagent, JT97-N002S)


NTRII 轉染試劑產品 (NTRII, #JT97-N002S) 3000/ml,具有高效能、低價位及高品質等特點。其價格為一般市售商品的1/3~1/5倍,因為適用於大部份的細胞株 (效能如下所示),所以廣受研究單位所使用。


 


更多參考資料如下所示:


分類:細胞轉染試劑 (T-Pro NTRII)


http://tw.myblog.yahoo.com/taiding2000/article?mid=209&prev=367&next=184&l=f&fid=16


 


3.Lentivirus病毒表現載體. Lentivirus病毒表現. Lentivirus病毒濃縮. Lentivirus病毒純化. Lentivirus力價定量。


 


更多參考資料如下所示:


分類:miRNA Lentivirus慢病毒表現、定量


http://tw.myblog.yahoo.com/taiding2000/article?mid=187&prev=191&next=179&l=f&fid=12


 


4. Cell immortalizing產品系列。


lenti-hTERT,p53,Myc,Rb,Ras V12, SV40 T antigen誘導細胞不杇。.


 


更多參考資料如下所示:


分類:間質幹細胞株、SV40病毒細胞不杇、初代細胞http://tw.myblog.yahoo.com/taiding2000/article?mid=333&next=200&l=f&fid=22


 


三、  細胞添加劑


 


1. 生長激素(Cytokines), 趨化因子(Chemokines), 生長因子(Growth Fectors), 荷爾蒙(Hormones), Viral Antigens和各式抗體(antibody)


ProSpec產品種類超過千種,產品涵蓋;生長激素(Cytokines), 趨化因子(Chemokines), 生長因子(Growth Fectors), 荷爾蒙(Hormones), Viral Antigens和各式抗體(antibody), 廣泛應用於研發、診斷試劑之開發、組織培養或無血清培養基之添加,癌症、細胞凋零、發育、內分泌、免疫、神經科學、蛋白質體、及幹細胞等相關領域的研究。


 


更多參考資料如下所示:


分類:ProSpec重組蛋白、單多株抗體 http://tw.myblog.yahoo.com/taiding2000/article?mid=255&prev=259&next=239&l=f&fid=27


 


2.      黴漿菌除去試劑MycoawayG398


其在市場上具有競爭力,其可移除超過70種以上Mycoplasma species。僅需要4次處理。1:1,000倍稀釋為治療。10,000倍稀釋為預防。


更多參考資料如下所示:


分類:分類:黴漿菌偵測套組、抑菌劑、抗生素添加http://tw.myblog.yahoo.com/taiding2000/article?mid=381&prev=382&next=346&l=f&fid=36


 


四、  細胞試驗 (細胞凋亡、細胞代謝分析、細胞氧化傷害分析、細胞增殖、細胞毒理分析、ELISA分析)


 


1.     細胞代謝分析


代謝質體學是基於功能性基因體學的觀念﹑對細胞進行全面檢測其體液或體組織中代謝中間物及代謝產物之組成﹒BioVison提供百樣代謝體(metabolism)研究試劑﹐方便您metabolism pathway研究﹒


 


更多參考資料如下所示:


分類:BIOVISION代謝分析(脂質、糖類、有機物)


http://tw.myblog.yahoo.com/taiding2000/article?mid=43&prev=231&l=f&fid=37


 


2.     Biovision細胞凋亡產品種類眾多。


如何選擇適合的Apoptosis assay kitBiovsion提供你最快的產品訊息。依樣品種類、觀察細胞的不同位置、研究的目的不同,細分有幾十種產品。


更多參考資料如下所示:


分類:Biovision 細胞凋亡、細胞增殖http://tw.myblog.yahoo.com/taiding2000/article?mid=236&prev=246&next=235&l=f&fid=30


 


3.     細胞氧化傷害 (comet assay)


彗星分析方法,可直接觀察單一細胞DNA損傷的程度。是一種快速、 簡易性、及高靈敏度檢測各別細胞DNA受損的方法(Singh et al., 1988) 應用電泳設備,即可進行分析


 


更多參考資料如下所示:


分類:DNA損害產品 、Comet assay彗星試驗http://tw.myblog.yahoo.com/taiding2000/article?mid=274&prev=349&next=258&l=f&fid=13


 


4.     細胞或上清液ELISA分析


更多參考資料如下所示:


分類:ELISA 酵素免疫分析套組


http://tw.myblog.yahoo.com/taiding2000/article?mid=172&prev=327&next=87&l=f&fid=39


 


5.     細胞增殖分析 (WST呈色分析或螢光分析套組)


安全性佳, 除了測量細胞增殖分析外, 可以再進行下游分析


 


更多參考資料如下所示:


分類:Biovision 細胞凋亡、細胞增殖http://tw.myblog.yahoo.com/taiding2000/article?mid=101&prev=102&l=f&fid=30


 


6.     細胞毒理分析 (LDH分析套組)













K311-400 



 LDH-Cytotoxicity Assay Kit  



 400 assays 



 K313-500 



 LDH-Cytotoxicity Assay Kit II  



 500 assays 



 


五、  細胞污染防治及黴漿菌污染偵測


 


1.      Mycoplasma 黴漿菌快速PCR鑑定。


應用PCR原理, 快速及高靈敏度2~ 5 f g/DNA的方法,偵測潛在性的mycoplasma污染。準確度高達95%以上。


 


更多參考資料如下所示:


分類:黴漿菌偵測套組、抑菌劑、抗生素添加


http://tw.myblog.yahoo.com/taiding2000/article?mid=46&prev=94&l=f&fid=36


 


2.      CO2 細胞培養箱或水浴槽T-Pro Aqua EZ Clean,去黴防菌抑菌劑。


可有效預防黴漿菌細菌之污染。


 


更多參考資料如下所示:


分類:黴漿菌偵測套組、抑菌劑、抗生素添加


http://tw.myblog.yahoo.com/taiding2000/article?mid=144&prev=346&next=94&l=f&fid=36


 


六、  粒腺體染劑、活細胞染劑


1. 粒線體染劑JC-1 (Catalog#1130-5),測量粒線體膜電位改變


 


更多參考資料如下所示:


分類:Biovision 細胞凋亡、細胞增殖http://tw.myblog.yahoo.com/taiding2000/article?mid=102&prev=103&next=101&l=f&fid=30


 


2. 活細胞染劑 “Calcein AM”


用於活細胞觀察及細胞侵入及轉移試劑.Calcein-AM是一種可對活細胞進行螢光標記的細胞染色試劑,Calcein-AM由於在Calcein(鈣黃綠素)的基礎上加強了疏水性,因此能夠輕易穿透活細胞膜。當其進入到細胞質後,酯酶會將其水解為Calcein(鈣黃綠素)留在細胞內,發出強綠色螢光。


 


更多參考資料如下所示:


分類:Biovision 細胞凋亡、細胞增殖http://tw.myblog.yahoo.com/taiding2000/article?mid=61&prev=91&l=f&fid=30


 


 


 


 

太鼎生物科技 發表在 痞客邦 留言(0) 人氣()


Stable Transfection


 


Stable Transfection (例如使用G418篩選)


 



篩選之前請先確定G418抗生素濃度: 


 


 


1.          由於每種細胞對G418抗生素的敏感性不同,而且不同的廠家生產的G418有效成分的比重不同,一般1g 的粉劑中有效的G418抗生素含量大約為0.722g


2.          G418抗生素是Neomycin的類似物,兩者都是通過抑制核糖體的功能和蛋白質的合成而殺死細胞的。但是Neomycin對真核細胞無作用而G418抗生素對細菌和真核細胞都起作用。在進行轉染時細胞膜受到影響,抗生素可能對細胞產生較大影響,加上G418抗生素有殺菌作用,所以有人主張轉轉染時不加其它抗生素。


3.          細胞密度(滿盤率)G418抗生素篩選結果的影響很大,一般篩選時細胞密度(滿盤率)不宜超過50%


4.          G418抗生素的活性不盡相同,所以在篩選之前,一定要確定G418抗生素的最佳篩選濃度。具體如下:將細胞稀釋到1000個細胞/ml,在100ug/ml~1mg/mlG418濃度範圍內進行篩選,選擇在10~14天內使細胞全部死亡的最低G418抗生素濃度調控來進行下一步的篩選試驗。


 


加藥時間和維持濃度:


 


1.          由於基因轉染到細胞內之後要一段時間才能表達出蛋白質。所以篩選不能太早;但是也不能太晚,因為轉染了外來基因的細胞代謝負荷較大,增值較慢,時間長了就會被沒有外來基因轉入的細胞所淹沒,最終導致篩選不出陽性,一般要在轉染24小時之後才開始加G418抗生素篩選。隨著細胞的代謝G418抗生素的濃度和活性都會下降,所以每3~5天都要更換一次含有G418抗生素的篩選液。篩選時G418抗生素濃度可以調控高中低三種進行篩選。


2.          G418抗生素的時機,主要是考慮插入到細胞基因模組的抗性基因是否已經得到表達。一般是轉染24小時後加入G418抗生素。


3.          在挑出單一細胞克隆株後就可以用固定G418抗生素濃度,一般是篩選時濃度的1/2
(
關於固定維持G418抗生素濃度,有人說細胞會出現對抗生素的抗性,應不斷提高其濃度。而且,如果你要挑選到幾個陽性克隆中較高表達的的話,可以調整G418抗生素的濃度。當然,抗性基因高表達的單一細胞克隆株,想表達的基因不一定就會跟著高表達。)


 


篩選時的培養液:


 


G418抗生素篩選約6天左右,細胞會大量死亡,培養盤中剩下的細胞寥寥無幾。這時會出現兩個問題:


1.          死亡的細胞會裂解釋放出有害物質,導致那些有G418抗生素表達的陽性細胞死亡,非選擇性死亡,因此要及時換液。


2.          培養盤中細胞數目很少,細胞之間的信號會變得很弱,也會導致陽性細胞的狀態不佳甚至死亡。這個時候需要一種特殊的培養液:例如你要轉染3T3細胞,先在3T3細胞密度(滿盤率)達到80%的時候,在換液時將培養過的舊培養液收集,使用過濾器無菌處理,和新鮮的培養液按11混合備用。在轉染後加G418抗生素篩選過程中就可以使用這種培養基。


3.          適當增加血清濃度(15%)


 


篩選時出現的問題及其解決辦法:


 


問題1:使用HeLa Cell篩選時,現在已經篩選3週,在6孔盤中已經長出一些細胞簇,想把它挑到96孔盤中,使用2ul胰蛋白酶消化,在消化過程中,胰蛋白酶擴散,使細胞散開和周圍的其它細胞簇融合在一起(我的細胞簇彼此距離的很近),造成幾個細胞簇被胰蛋白酶融合在一起,那樣根本沒有辦法做下去了,該怎麼辦?
a
、可以減少胰蛋白酶使用量;且胰蛋白酶在加入之前要用37 恆溫箱中預熱,並且使用PBS潤洗細胞層,以減少可能殘存的血清的影響;或是改使用(ACCUTASE)

b
、加入胰蛋白酶後,稍過幾秒鐘就可以吸走消化液了,不用吸乾淨,然後放到37 恆溫箱中繼續作用1分鐘,這時,胰蛋白酶消化細胞可以繼續作用的,又可避免胰蛋白酶擴散。
c
、顯微鏡下觀察細胞完全鬆散開,就可以在你感興趣的局部加培養基,並吸走你的可隆了呀
d
、具體消化的時間需注意,如果在步驟b中顯微鏡下見細胞尚未完全鬆散開,可以再重複的,直到鬆散開,能夠被吸走為止。
e
、或在100mm dish中挑克隆,細胞分的稀一點。
f
、也可把細胞全部消化下來,在96孔盤中逐步稀釋獲得單一細胞。


問題2:篩選成功的機率;只要有抗性加壓篩選,挑選到的機率還是比較大的。能挑到穩定表達的機率一般認為大約有70~80%,但是想挑到表達量高且能穩定表達的,並不容易。這個可能是在染色體上,合適的整合位點太少的緣故。
問題3:單一陽性細胞克隆化的時機和個數:加抗生素篩選時,一般會等到確認已轉染細胞達到70%以上時,再做有限稀釋法,使克隆出陽性細胞,同時要保持適當的抗生素濃度,並注意突變和污染。若細胞生長(滿盤率)如有40%以上,就可以做有限稀釋了,一般篩選56個克隆是有需要的,但保險起見,多篩幾個吧。從單一陽性細胞克隆化時開始,就要增加對細胞營養及血清含量和生長因子。

太鼎生物科技 發表在 痞客邦 留言(0) 人氣()


Transfection Reagent (in vivo)


 



說明:
T-Pro G – Fect (體內)轉殖試劑是帶有陽離子的無菌溶液,因其帶正電可與DNA形成穩定的複合物。在生物體內進行基因轉殖的過程中,此複合物的型態可保護DNA不受降解,同時使轉殖的過程更容易。傳送DNA的過程可通過各種途徑,包括靜脈注射,腹腔,腫瘤內注射等,而試劑在這樣的使用過程中並未檢測到發炎反應的產生。


 


試劑提供:
Reagent A或使用無菌 5W/ V葡萄糖溶液代替做為稀釋的DNA之用。



一般注意事項:
DNA的品質要求
 DNA
的品質是轉殖是否成功的重要關鍵。建議
A260/A280比值1.8或者更高。同時DNA應該是無菌的,且不具任何污染物,如內毒素。


選擇啟動子(promoter)
 在標的組織 /器官中,高目標基因表現量取決於啟動子與目標基因的選用。巨細胞病毒(CMV)啟動子是最常使用於各類細胞中。但有些研究也常使用LTRSV40)或(RSV)作為啟動子(promoter)





轉殖試劑 / DNA比率:
轉殖試劑的使用量取決於DNA的量,轉殖基因使用量和被轉殖動物的比率可先參照(1)。再依據結果加以調整取得最佳效果。


 
DNA注射建議量:
DNA注射量,取決於所使用實驗動物的種類和注射方式(見表 1),以及標的組織或器官。
為了避免T-Pro G - Fectin vivo/ DNA複合物沉澱,建議注射DNA的最終濃度不應超過0.5μg/μl

太鼎生物科技 發表在 痞客邦 留言(0) 人氣()

T-Pro Mounting Reagent or T-Pro Mounting Long Reagent 是一種高效性抗退色試劑,可使用於免疫螢光染色樣品之觀察。此一高效性抗退色試劑可幫助螢光樣品抵抗於螢光顯微鏡下觀察時所產生之光漂白現象。


本產品之使用能適用於各種螢光染劑或抗體。並且使用本產品不但能使螢光樣品之觀察時間增長進而增加實驗成功率更可增加樣品保存期限至最多1星期。 


 


特性:


使用前請將“T-Pro Mounting Reagent or T-Pro Mounting Long Reagent 回溫至室溫。


在使用“T-Pro Mounting Reagent or T-Pro Mounting Long Reagent 前請盡量去除樣品上殘留之液體。


• “T-Pro Mounting Reagent or T-Pro Mounting Long Reagent 適用於大多數樣品於差異干擾差、相位差或螢光顯微鏡下觀察。


 


操作:


1.回溫試劑:


“T-Pro Mounting Reagent or T-Pro Mounting Long Reagent ”4oC 冰箱中取出並回溫至室溫備用。請勿將“T-Pro Mounting Reagent or T-Pro Mounting Long Reagent ”以溫水或其他加熱方式回溫,以免破壞產品穩定性。


2.使用Apply T-Pro Mounting Reagent or T-Pro Mounting Long Reagent


在使用“T-Pro Mounting Reagent or T-Pro Mounting Long Reagent ”前請將樣品玻片上之殘留液體以拭鏡紙由玻片邊緣輕輕去除並在玻片上滴上一滴“T-Pro Mounting Reagent or T-Pro Mounting Long Reagent ”10-20ul)。將樣品蓋玻片以45度角,緩慢的蓋在含有“T-Pro Mounting Reagent or T-Pro Mounting Long Reagent ”之載玻片上,其間請避免氣泡之產生。 


3.上機前準備


樣品玻片完成步驟二之製備後,請避光保存於4oC冰箱中並請注意保存時表面之平坦。保存期限則因樣品的不同而有所差異,自數小時至隔夜不等,同時也受到儲存環境中溫濕度的影響。如需較長時間保存,建議將樣品玻片以指甲油封片,並避光保存於4oC冰箱中。必要時,保存盒內可放置乾燥劑以維持儲存環境之乾燥。


 



1. T-Pro Mounting Long Reagent I 有幫助螢光分子抵抗光漂白之效果。圖1.a NIH 3T3細胞以4%福馬林固定後再以Fluorescein-phalloidin對細胞骨架進行染色。 圖中影像乃是將染色完成之樣品置於T-Pro Mounting Long Reagent IPBS中,並在封片後以100瓦汞燈對樣品進行不同時間之照射以產生光漂白之效果,再以63x/1.4 NA 油鏡及16-bit 單色 CCD攝影機在相同條件下進行拍攝。在所得影像中可清楚得知於PBS對照組中,在螢光照射120秒以後,絕大部分之螢光分子因光漂白而無法呈色,而在實驗組中發現T-Pro Mounting Long Reagent I能幫助螢光分子抵抗光漂白,直至螢光照射7分鐘後,大部分之螢光分子才因光漂白而無法呈色,因此說明了T-Pro Mounting Long Reagent I有幫助螢光分子抵抗光漂白之效果。並將兩組實驗在不同時間所取得之影像,以軟體對螢光衰減率進行分析後所做之圖。


 



下圖、以T-Pro Mounting Long Reagent I with DAPIAlexa Fluo 488 phalloidinAlexa Fluo 594 wheat germ agglutininNIH 3T3細胞進行染色之成果。





太鼎生物科技 發表在 痞客邦 留言(2) 人氣()




Transfection 如何經費?  


 


    


NTRII 轉染試劑產品 (NTRII, #JT97-N002S) 3000元/ml,具有高效能、低價位及高品質等特點。其價格為一般市售商品的1/3~1/5倍,因為適用於大部份的細胞株 (效能如下所示),所以廣受研究單位所使用。其使用的比例較PJ牌為低,用量很省,一般的細胞株其用量為1ug DNA: 2ul NTRII。因其毒性比Liposome低,因此更適用於primary cell。建議客戶使用便宜的NTRII來轉染細胞株。遇見難以轉染的細胞株,例如巨噬細胞、肌肉細胞、心肌細胞,再用高價位的轉染試劑,或是病毒等方式來進行。此產品在中研院、國衛院、台大等單位都一直使用。若須試用品100ul試用,請洽各區域業務。   


LentiVirus病毒蛋白於293T細胞株的產製,應用傳統方式,以10公分培養基,須要10ugDNA的濃度。中研院客戶使用NTRII轉染包裝載體於293T細胞株,每10分培養基,只須4ug的DNA用量。應用NTRII轉染試劑來表現Lentivirus蛋白,其方法簡單、DNA用量少、具有高病毒蛋白的表現量。 


                                  


【轉染試劑產品介紹】


T-Pro Non-liposome transfection Reagent II 對各種不同的細胞株進行轉染測試,其效果都很好。其中包括肝癌細胞HepG2Huh7; 乳癌細胞株MCF7; 神經細胞株N 2A ;  HEK293, 293T, 293E, BHK, CHO懸浮或貼附, COS1, COS7, HeLa, Vero, L929, NIH 3T3, Human Foreskin Fibroblasts (HFF) primary cells, Bovine Aorta Endothelial cells(BAEC), Sf9Sf21昆虫等常用細胞株。


NTRII產品特點】


- 不具細胞毒性 ----更適用於Primary cell


- 在廣泛的細胞株測試中皆具有極佳的轉染效果 -----好轉的細胞株,就用便宜的轉染試劑。


- 可在有血清環境下使用具有極佳的轉染效果 ----不須6小時後更換medium


- 可產生大量的重組蛋白 ------Lentivirus病毒產製10公分dish只須4ug DNA: 10ul NTRII


- 操作步驟簡單快速 ----Lipofetamin2000簡單


- 無論對於貼附性細胞或是懸浮性細胞皆具有極佳的轉染效果


【操作指南】


哺乳動物細胞基因轉染的操作流程:


 1.對於貼附型細胞


細胞之數量(見表1):細胞應於轉染前1824小時前培養,以確保細胞數量可以在轉染前密度達約40~90%之單層


細胞狀態並於轉染前0.5-2小時前將細胞培養液更新。在轉染時,請使用含血清之細胞培養液,但於某些特殊情形下,


可使用不含抗生素之無血清培養液或血清少於5%之培養液,可改善重組蛋白的表現量或有助於轉染之效果。


T-Pro Non-liposome transfection Reagent II -DNA複合物的製備和轉染操作步驟


1. 對於不同的細胞型態,T-Pro Non-liposome transfection Reagent II (μl)DNA (μg)較佳的比例為11~41


在一開始時,我們採用的比例為21且通常可以得到滿意的轉染效率且不具細胞毒性。在某些特殊狀況下,可嘗試改變混


和比例以達到最佳轉染效果。為了確保混合物的最佳效果,我們建議採用不含血清及抗生素之細胞培養液或HBS buffer


(pH7.4 150m M NaCl, 20m M HEPES) DNAT-Pro Non-liposome transfection Reagent II進行稀釋。以下轉染操作


流程適用於24孔細胞培養盤,其他培養盤之用量可依據其體積稍作調整,在表2有提及其他形式培養皿的轉染條件可供參考。


 


2. 24孔細胞培養盤每孔為例,將1μgDNA30μl (不含血清及抗生素之細胞培養液)HBS buffer


(pH7.4 150mM NaCl, 20mM HEPES)混合。


3. 2μlT-Pro Non-liposome transfection Reagent II 加入步驟2DNA之混合物進行混合,經混合後於室溫下靜置15分鐘。


4. 再將步驟3之混合物均勻滴於盤內(含有細胞及血清培養液),並以前後左右搖晃的方式使其均勻混合。


5. 將轉染後之細胞置於37℃ 5 CO2的培養箱中培養。


6. 1848小時後確認轉染及表現效率。


注意:若需轉染其他多孔盤或培養皿時,請將所需體積等比例放大或縮小。


2.對於懸浮細胞


細胞數量:懸浮細胞於轉染當天將細胞置換於新的培養液中並將其數量控制在0.5-1× 106 c ells/ml間。


T-Pro Non-liposome transfection Reagent II -DNA複合物的製備和轉染操作步驟


1. 對於不同的細胞型態,T-Pro Non-liposome transfection Reagent II (μl)DNA (μg)較佳的比例為11~41。在一開始時,我們採用的比例為21且通常可以得到滿意的轉染效率且不具細胞毒性。在某些特殊狀況下,可嘗試改變混和比例以達到最佳轉染效果。為了確保混合物的最佳效果,我們建議採用


 不含血清及抗生素之細胞培養液或HBS buffer(pH7.4 150m M NaCl, 20m M HEPES)DNAT-Pro Non-liposome transfection Reagent II進行稀釋。以下轉染操作流程適用於24孔細胞培養盤,其他培養盤之用量可依據其體積稍作調整,在表2有提及其他形式培養皿的轉染條件可供參考。


2. 24孔細胞培養盤每孔為例,將1μgDNA60μl (不含血清及抗生素之細胞培養液)HBS buffer(pH7.4 150mM NaCl, 20mM HEPES)混合。


3. 2μlT-Pro Non-liposome transfection Reagent II 加入步驟2DNA之混合物進行混合,經混合後於室溫下靜置15分鐘。


4. 再將步驟3之混合物均勻滴於盤內(含有細胞及血清培養液),並以前後左右搖晃的方式使其均勻混合。


5. 將轉染後之細胞置於37℃ 5 CO2的培養箱中培養。


6. 1848小時後確認轉染及表現效率。


注意:若需轉染其他多孔盤或培養皿時,請將所需體積等比例放大或縮小。\


 


T-Pro Non-liposome transfection Reagent II -siRNA複合物的製備和轉染操作步驟


1. 對於不同的細胞型態,T-Pro Non-liposome transfection Reagent II (μL)siRNA (μg)較佳的比例為11~41在一開始時,我們採用的比例為31,且通常可以得到滿意的轉染效率且不具細胞毒性。在某些特殊狀況下,可嘗試改變混和比例以達到最佳轉染效果。為了確保混合物的最佳效果,我們建議採用不含血清及抗生素之細胞培養液或HBS buffer(pH7.4 150m M NaCl, 20m M HEPES)T-Pro Non-liposome transfection Reagent   II進行稀釋。以下轉染操作流程適用於24孔細胞培養盤,其他培養盤之用量可依據其體積稍作調整,在表2有提及其他形式培養皿的轉染條件可供參考。


2. 24孔細胞培養盤每孔為例,取30μl (不含血清及抗生素)細胞培養液或HBS buffer(pH7.4 150mM NaCl, 20mM HEPES)


3. siRNA 5~50 nmol 加入到步驟2中均勻混合。


4. T-Pro Non-liposome transfection Reagent II 1~5μl 加入到步驟3siRNA之混合物均勻混合,經混合後於室溫下靜置15分鐘。


5. 將步驟4之混合物取均勻滴於盤內(含有細胞及血清培養液),並以前後左右搖晃的方式使其均勻混合。


6. 將轉染後之細胞置於37℃ 5 CO2的培養箱中培養。


7. 1848小時後確認轉染及表現效率。


注意:若需轉染其他多孔盤或培養皿時,請將所需體積等比例放大或縮小。


注意:siRNA溶液濃度,需在1μM 以上使用。


更多參考內容:


 


WWW.BIOPIONEER.COM.TW


 


太鼎生物科技有限公司(02)86609496


 



業務專員: 許 虹宜Hung-Yi Hsu       


電話: 0920312382




 

太鼎生物科技 發表在 痞客邦 留言(0) 人氣()

                         RNAi專用的轉染試劑


                        RNAifectin™ Transfection Reagent, G073



 


【太鼎生物科技有限公司02-86609496abmgood台灣總代理


 










Cat.No.:G073
Quantity:1.0ml (100-500 transfections)





















Description



Escpecially developed for the efficient transfection of eukaryotic cells with RNAi oligo's. This transfection reagent is perfect for any of your RNAi transfection needs.



Application



RNAi knockdown experiments in a wide variety of cell lines (HeLa, HEK-293 etc). Highly specific and non-toxic transfection.



Protocol





Storage



Store at 4ºC. Do not freeze.



Notes



This product is distributed for laboratory research only. Caution: Not for diagnostic use .



產品介紹 Description


ANAifectin™ is a transfection reagent specially formulated with multiple cationic polymers. It is suitable for the transfection of RNAi oligoes into cultured eukaryotic cells.


RNAi轉染操件手冊:


These conditions are recommended as guidelinesonly. A 6-well or 35mm dish is adequate for most applications, but larger vessels are sometimes required. In that case, consult table 1.



1. In a 6-well plate, seed cells at a density of 1-3x10⁵ per well in 2.0ml of the appropriate
growth medium (with serum if cells are cultured in presence of serum). Incubate the cells at 37°C until cells are 60-90% confluent. This will usually take 18-24 hours, depending on cell types. Since transfection efficiency is sensitive to culture confluence, it is important to maintain a standard seeding protocol from experiment to experiment


2. Prepare the following solutions in sterile 12x75 mm or microcentrifuge tubes: Solution A: For each transfection, dilute 1-3μg of RNAi oligoes into 125μl serum-free
medium. Solution B: For each transfection, dilute 4-10μl of
RNAifectin™ reagent into 125μl of serum-free medium. (Vortex RNAifectin™ reagent before use)


3. Combine the solutions, mix thoroughly and incubate at room temperature for 20 mins to
allow DNA-lipsome complexes to form.


4. Remove growth medium from the cells and add 800μl of serum-free medium to the cells.


5. Add the RNAifectin™-oligo complexes to thecells, mix gently to ensure uniform distribution
and incubate at 37°C for 2-24 hours. We recommend starting with 5 hours.


6. Remove the transfection solution and add 2.0ml of the appropriate complete growth medium(with serum) to each well. Incubate cells at 37°Cfor a total of 24-72 hours.


7. Assay cell extracts for gene activity 24-72 hours a􀄞er the start of transfection depending on the cell types and promoter activity.


8. A similar procedure can be used to transfect an RNAi vector for stable expression. At 72 hours a􀄞er transfection, split the cells at a ratio of 1:10 into the selective medium for the marker gene transfected.


【太鼎生物科技有限公司02-86609496abmgood台灣總代理



業務專員: 許 虹宜Hung-Yi Hsu       


電話: 0920312382




太鼎生物科技 發表在 痞客邦 留言(0) 人氣()

1 2