抗體純化套組 Abselect kit
移除溶液中BSA、Glycine、Tris、Azide等殘留分子，純化後溶液，可直接應用於奈米金抗體標誌套組 (Innova gold,#230-0010, innova biosciences)，奈米金抗體標誌, 相關資料來源: http://tw.myblog.yahoo.com/taiding2000/article?mid=579&prev=615&next=576&l=f&fid=19
BSA、Gelatin、Glycine移除--AbSelect 抗體-----純化系統
適合應用: 市售抗體。預移除溶液中BSA、Glycine、Tris、Azide等殘留分子
純化體積100ul至500ul
AbSelect抗體純化套組 (860-0010), 特別針對市售抗體保存液內, 含有BSA, Glycine. Tris. Azide等amino group干擾物質殘留。純化後的抗體, 可直接應用於奈米金抗體標誌套組 (Innova gold,#230-0010, innova biosciences)及抗體標誌試劑 (lightning-link一個步驟完成抗體標誌試驗)。
不須經由dialysis等純化步驟。抗體的體積可經由100ul至0.5ml， 每次純化, 可以純化>500ug抗體量。
Ps. 蛋白質濃縮及buffer exchange之動作, 無法有效移除抗體內的小分子, 須經由Abselect 抗體純化系統來執行。
AbSelect訂購貨號
1個純化套組, 貨號#860-0005
3個純化套組, 貨號#860-0010
AbSelect套組內容物
Lyophilised AbSelect Protein A resin
1 vial of 10x Binding Buffer
1 vial of Wash Buffer
1 vial of Elution Buffer
1 vial of Neutralization Buffer
1 or 3 spin cartridge/collecting tube assemblies
4 or 12 additional collecting tubes
AbSelect抗體純化流程
Step 1 取Wash buffer重組Protein A resin
Step 2 將重組好的Resin, 移至離心管, 並spin down
Step 3 抗體加入10x Binding Buffer後,移入管柱中反應1個小時
Step 4 Wash resin.
Step 5 Elute 及neutralize約化的antibody.
Step 6 確認elute的抗體含量, 並以蛋白質定量.
AbSelect Antibody Purification System
Commercially available antibodies often contain substances (e.g. BSA, glycine, tris, azide) that interfere in labeling reactions. The AbSelect™ Purification Kit quickly removes these contaminants. It can also be used to purify antibodies from crude samples such as ascites fluid or immune serum.
The antibody to be purified or cleaned up ideally is in a volume of 100ul to 0.5ml. Up to 500µg of antibody can be purified in each run. The AbSelect method involves capture of the antibody on protein A resin and the removal of unwanted substances by a simple wash procedure. The purified product is then eluted and neutralized.
The components of the AbSelect kit are fully compatible with the Lightning-Link™ antibody labeling system (available separately), which allows the purified antibody to be rapidly conjugated with a wide variety of labels.
腹水、血清純化抗體 AbSelect 血清抗體------純化系統
適合應用: 腹水及血清產製之抗體。純化來自培養液、腹水及血清之抗體，移除殘留之amino group。純化後之抗體，可直接進行奈米金抗體標誌套組 (Innova gold,#230-0010, innova biosciences)及抗體螢光或HRP標誌(lightning_link)。
AbSelect Serum Antibody Purification System
Antibodies generated from ascites fluid and serum are often supplied as crude formulations. Both ascites fluid and serum contain many substances that interfere with antibody labeling reactions. Successful labeling therefore requires antibodies to be purified prior to labeling. The AbSelect Serum system is a fast and simple method to purify antibodies from these types of media.
AbSelect Serum is prepared by coupling highly purified protein A to agarose beads which can be used to purify IgG fractions from ascites fluid and serum. The method involves the capture of the antibody on the AbSelect Serum resin and the removal of unwanted substances using a simple wash procedure. The purified product is then eluted and neutralized.
The components of the AbSelect Serum purification system are fully compatible with the Lightning-Link™ antibody labeling kits (available separately), which allows the purified antibody to be immediately labeled with an enzyme, fluorescent protein or fluorescent dye, with a hands-on time of under 30 seconds.
腹水、血清純化抗體 AbSelect TCS組織培養液抗體------純化系統
適合應用: 融合瘤組織培養液(TCS)產製之抗體。純化來單株細胞產製之抗體，移除培養液中殘留之amino group。純化後之抗體，可直接進行奈米金抗體標誌套組 (Innova gold,#230-0010, innova biosciences)及抗體螢光或HRP標誌(lightning_link)。
AbSelect TCS Antibody Purification System
Antibodies are often generated from hybridoma cell lines and supplied in tissue culture supernatant (TCS). TCS itself contains many substances that interfere with antibody labeling reactions. Successful labeling therefore requires antibodies to be purified prior to labeling. The AbSelect TCS system is a fast and simple method to purify these antibodies.
AbSelect TCS is prepared by coupling highly purified protein A to agarose beads which can be used to purify IgG fractions from hybridoma supernatants. The method involves the capture of the antibody on the AbSelect TCS resin and the removal of unwanted substances using a simple wash procedure. The purified product is then eluted and neutralized.
The components of the AbSelect TCS are fully compatible with the Lightning-Link™ antibody labeling kits (available separately), which allows the purified antibody to be immediately labeled with an enzyme, fluorescent protein or fluorescent dye, with a hands-on time of under 30 seconds.
抗體濃縮及Clean up系統
適合應用: 低濃度抗體之純化及濃縮。干擾酵素型HRP、Biotin、streptavidin及FITC螢光等之抗體標誌反應。移除溶液中殘留之amino group。純化後之抗體，可直接進行奈米金抗體標誌套組 (Innova gold,#230-0010, innova biosciences)及抗體螢光或HRP標誌(lightning_link)。
抗體濃縮及Clean up流程
Step 1 加入 0.5ml 以上的抗體量至 spin cartridge.
Step 2 15,000g離心 1 to 3 分, 使抗體量降低至100ul
Step 3 吸掉離心管的溶液
Step 4 加入 400μl conjugation buffer至spin cartridge中的抗體
Step 5 15,000g離心 1 to 3 分, 使抗體量降低至100ul
Step 6吸掉離心管的溶液.
Step 7 重覆1-6步驟, 置換抗體溶液。
Step 8 由spin cartridge中回收純心後的抗體
Antibody Concentration and Cleanup Kit
Antibodies are sometimes only available at low concentrations and often contain low molecular weight substances that interfere in labeling reactions with enzymes, biotin, streptavidin and fluorophores. The Antibody Concentration and Clean Up Kit allows for the quick and easy concentration of antibodies and proteins. The kit can also be used to reduce the concentration of many unwanted additives often found in antibody formulations such as azide, glycine or tris.
The antibody clean up kit method utilizes a simple spin column to easily and quickly remove excess buffer from the antibody thereby providing a more concentrated antibody solution. The clean up kit also allows the experimenter to perform a simple buffer exchange to transfer the antibody into a more favorable conjugation buffer.
DIY抗體標誌螢光(FITC,PE,Cy3,Cy5,Cy7 dye)
注意事項
請確認抗體的保存條件。傳統市售的抗體標誌套組，在使用前皆需dialysis純化，將抗體保存的物質移除。Lightning-link抗體標誌套組在大部份的保存條件下，不需要再dialysis的步驟，例如一般salts (e.g. NaCl)、sugars(e.g. sucrose)、及chelators (e.g. EDTA)不會影響標誌的活性。大部的市售的抗體保存在1%BSA、0.5%Azide及 0.5% Gelatin 輕微或幾乎不會影響標誌的活性。在少數情況下，若是您購買的抗體保存在amino acids(e.g. glycine)、blockers(e.g. ethanolamine)及thiols(DTT, mercaptoethanol)情況下，記得純化您的抗體，(abselect 抗體純化套組) 避免抑制您的標誌活性了!
【供應廠商】太鼎生物科技有限公司
【連絡人】許虹宜0920-312382
【連絡信箱】taiding.biotech@msa.hinet.net
【台北公司】02-86609496 【台中公司】0935-229803 陳俊豪
【公司網址】 www.biopioneer.com.tw
【原廠網址】http://www.innovabiosciences.com
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