The one-step staining procedure takes only 10 minutes. Annexin V-FITC conjugate is specially engineered to produce enhanced fluorescence signal and photo stability. The Annexin V-FITC kit includes Annexin V-FITC for detecting apoptosis and also propidium iodide (PI) for detecting necrosis. Thus, Apoptosis and necrosis can be differentiated. Detection can be analyzed by flow cytometry (Ex/Em=488nm/530nm) or by fluorescence microscopy using a dual filter set for FITC and rhodamine. The Annexin VFITC Plus kit includes cytox green dye (instead of PI) which stains necrotic cells with a higher level of green fluorescence than FITC does. Thus, both apoptosis and necrosis can be detected using FL1 channel.
The antiserum was produced against synthesized non-phosphopeptide derived from human NF-κB p65 around the phosphorylation site of serine 276 (R-P-SP-D-R).
Species Reactivity
Human, Mouse and Rat
Specificity
NF-κB p65 (Ab-276) Antibody detects endogenous levels of total NF-κB p65 protein.
Tested Applications
WB and IHC
Application Notes
WB: 1:500~1:1000 IHC: 1:50~1:100
Raised In
Rabbit
Clonality
N/A
Purity
The antibody was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen.
Storage Buffer
phosphate buffered saline (without Mg2+ and Ca2+), pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.
Formulation
Liquid
Concentration
1ug/ul
Storage
Store at -20°C/1 year
References
Baeuerle P A, et al. (1994) Annu Rev Immunol. 12:141-179.
Baeuerle P A, et al. (1996) Cell 87:13-20.
Haskill S, et al. (1991) Cell 65:1281-1289.
Thompson J E, et al. (1995) Cell 80: 573-582.
Caution
NF-kB p65 (Ab-276) is for research use only and is not intended for therapeutic or diagnostic applications. Please contact a technical service representative for more information.
【NFkb產品介紹】 NFkB was identified as a sequence specific transcriptional activator that binds to the intronic enhancer of kappa light chain gene in B lymphocytes. NFkB is a heterodimer that consists of a 50 kDa DNA binding subunit (p50) and a 65 kDa transactivation subunit (p65/RelA). Both of these subunits exhibit sequence homology to the protooncogene c-Rel. The p50/p65 heterodimer remains in the cytosol in an inactive form as a complex with its inhibitor, IkB. Upon stimulation of cells by a wide variety of stimuli such as lipopolysaccharide (LPS), pro-inflammatory cytokines, and viral infection. IkB is phosphorylated and degraded by proteosome. The active NFkB heterodimer is translocated into the nucleus and induces gene expression.
Kit Summary: • Detection method- Absorbance (570 nm) or Fluorescence (Ex/Em=535/587) • Species reactivity- Mammalian • Application- The kit has a detection limit of 0.1 mU per well via colorimetric or 0.01 mU per well via fluorometric method, based on our unit definition.
Features & Benefits: • Simple procedure • Fast and convenient • The assay is simple, direct, highly sensitive and high-throughput adaptable
Kit Components: • Assay Buffer • OxiRed™ Probe (in DMSO) • H2O2 Substrate (0.88 M) • HRP Positive Control
Description: Peroxidases (EC number 1.11.1.x) are a large family of enzymes that typically catalyze a reaction of the form: ROOR' + electron donor (2 e-) + 2H+ → ROH + R'OH. For many of these enzymes the optimal substrate is hydrogen peroxide, but others are more active with organic hydroperoxides such as lipid peroxides. Peroxidases can contain a heme cofactor in their active sites, or alternately redox-active cysteine or selenocysteine residues. BioVision’s Peroxidase Assay Kit provides a convenient colorimetric and fluorometric means to measure the peroxidase activity in biological samples. In the presence of Peroxidase, the OxiRed Probe reacts with H2O2 in a 1:1 stoichiometry to produce the red-fluorescent oxidation product, resorufin. The resorufin is quantified by colorimetric (lmax = 570nm) or fluorometric methods (Ex/Em = 535/587 nm). The assay is simple, direct, highly sensitive and high throughput-ready. The detection limit is 0.1 mU per well via colorimetric or 0.01 mU per well via fluorometric method, based on our unit definition.
Storage Conditions: -20°C
Shipping Conditions: gel pack
USAGE: For Research Use Only! Not For Use in Humans.
Kit Summary: • Detection method- Absorbance (570 nm) • Sample type- Cell and Tissue lysates, culture media, urine, plasma and serum, as well as many other biological fluids • Species reactivity- Mammalian • Applications- Assay measuring either the combination of both small molecule and protein antioxidants or small molecules antioxidant alone in the presence of our proprietary Protein Mask.
Features & Benefits: • Simple procedure; takes less than 2 hours • Fast and convenient
Kit components: • Cu²ᶧ Reagent • Assay Diluent • Protein Mask • Trolox Standard
Description: Antioxidants play an important role in preventing the formation of and scavenging of free radicals and other potentially toxic oxidizing species. There are three categories of antioxidant species: enzyme systems (GSH reductase, catalase, peroxidase, etc.), small molecules (ascorbate, uric acid, GSH, vitamin E, etc.) and proteins (albumin, transferrin, etc.). Different antioxidants vary in their reducing power. Trolox is used to standardize antioxidants, with all other antioxidants being measured in Trolox equivalents. Measurement of the combined nonenzymatic antioxidant capacity of biological fluids and other samples provides an indication of the overall capability to counteract reactive oxygen species (ROS), resist oxidative damage and combat oxidative stress-related diseases. In some cases, the antioxidant contribution of proteins is desired whereas in other cases only the contribution of the small molecule antioxidants is needed. BioVision developed the TAC Assay Kit, which can measure either the combination of both small molecule antioxidants and proteins or small molecules alone in the presence of our proprietary Protein Mask. Cu²ᶧ ion is converted to Cu+ by both small molecule and protein. The Protein Mask prevents Cu²ᶧ reduction by protein, enabling the analysis of only the small molecule antioxidants. The reduced Cu+ ion is chelated with a colorimetric probe giving a broad absorbance peak around 570 nm, proportional to the total antioxidant capacity.
Storage Conditions: -20°C
Shipping Conditions: gel pack
USAGE: For Research Use Only! Not For Use in Humans
K274 :: Total Antioxidant Capacity (TAC) Assay Kit
• Herspring KF et al (2008) J Appl Physiol; 105: 1889-1896.
Kit Summary: • Detection method- Absorbance (540 nm) • Sample type- Cell and Tissue lysates, culture media, urine, plasma and serum, as well as many other biological fluids • Species reactivity- Mammalian • Applications- BioVision’s Nitric Oxide Colorimetric Assay Kit provides an accurate, convenient measure of total nitrate/nitrite in a simple two-step process.
Features & Benefits: • Simple two-step procedure; takes only ~1-1.5 hours • Fast and convenient • BioVision’s Nitric Oxide Colorimetric Assay Kit provides an accurate, convenient measure of total nitrate/nitrite.
Description: Nitric oxide (NO) plays an important role in neurotransmission, vascular regulation, immune response and apoptosis. NO is rapidly oxidized to nitrite and nitrate which are used to quantitate NO production. BioVision’s Nitric Oxide Colorimetric Assay Kit provides an accurate, convenient measure of total nitrate/nitrite in a simple two-step process. The first step converts nitrate to nitrite utilizing nitrate reductase. The second step uses Griess Reagents to convert nitrite to a deep purple azo compound. The amount of the azochromophore accurately reflects nitric oxide amount in samples. The detection limit of the assay is approximately 0.1 nmole nitrite/well, or 1 µM.
Storage Conditions: -20°C
Shipping Conditions: gel pack
USAGE: For Research Use Only! Not For Use in Humans
K262 :: Nitric Oxide Colorimetric Assay Kit
• Atochin, D. et al. Soluble Guanylate Cyclase a• 1β1 Limits Stoke Size and Attenuates Neurological Injury. Stroke, 2010; 41: 1815-1819.
• Caretti A et al (2008) Experimental Biology and Medicine 233: 1222-1230.
• Kim, H. W. et. al. Long-term blockade of vascular endothelial growth factor receptor-2 aggravates the diabetic renal dysfunction associated with inactivation of the Akt/eNOS-NO axis. Nephrol. Dial. Transplant., Apr 2011; 26: 1173 - 1188.
Bacterial luciferase uses the substrate bacterial luciferin which is a different molecule from D-luciferin. The plasmid encoding for bacterial luciferase also encodes for the substrate which is then synthesized by the cell, eliminating the neeed for exogenous substrate administration.
Renilla and Gaussia luciferase uses the substrate coelenterazine, again different from D-luciferin.
Firefly or clickbeetle luciferase catalyzes the oxidization of D-luciferin to oxyluciferin in the presence of ATP, Mg2+, and oxygen with production of a yellow-green light, shifting to red light in vivo at 37oC. Since ATP is required as a co-factor, Firefly luciferase can be used as an indicator of the presence of energy or “life” and function as a life-death stain.
Methods:
Dissolution
D-luciferin for in vitro use:
Dissolve 1g of D-luciferin in 33.3ml sterile water to make a 30 mg/ml * stock solution.
Mix gently, syringe filter (0.2 um), aliquot, purge with nitrogen or argon (inert gas prevents oxidation) and freeze under at -80°C for future use (up to 1 year), protect from light.
Thaw and keep on ice, protect from light. If the luciferin does not dissolve, briefly bring to temperature in a waterbath and gently mix.Dilute stock at 1:200 in complete tissue culture medium to 150ug/ml . incubate 5 min prior to imaging.
Dispose at end of day.
D-luciferin for in vivo use:
Dissolve 1g of D-luciferin in 66.6 ml in DPBS, w/o Mg2+ and Ca2+ to make a 15mg/ml solution*. Mix gently, syringe filter (0.2 um), aliquot, purge with nitrogen or argon (inert gas prevents oxidation) and freeze at -80°C for future use (up to 1 year), protect from light.
Thaw and keep on ice, protect from light. If the luciferin does not dissolve, briefly bring to temperature in a waterbath and gently mix. Dispose at end of day.
Inject at 150mg/kg, thus 200ul (3mg) for a 20 g mouse. Image in plateau phase (see kinetics curve).
1 g is sufficient for about 300 mice.
* D-luciferin can be dissolved up to a concentration of 50mg/ml or 178mM, it precipitates out at 60mg/ml. Na+ salt dissolves better than K+ salt, to 100mg/ml.
Injection routes:
Intraperitoneal (IP) injection:
injection of the substrate in the intraperitoneal cavity. A dose of 150mg/kg is recommended. Inject 200ul (of a 15mg/ml solution) for a 20 g mouse. (The maximum IP volume for a 20 g mouse is 1ml). Use a 25 G needle, 1 cc syringe. Take mouse in one hand, restrain the mouse belly up, hold the scruff of the neck between thumb and index and clamp the tail with the other fingers, point the head down (intestinal organs sag cranially). Inject half the dose in the lower left quadrant and the other half in the lower right quadrant for the most consistent results. Insert needle 5mm, bevel side up at a 30o angle.
Intravenous (IV) injection:
Intravenous injections in mice are routinely performed in the tailvein or retroorbital plexus. Even though an intravenous injection is a bolus injection, the same dose of 150mg/kg is recommended. Keep in mind that the maximal volume to inject IV is 10% of the blood volume. A 20g mouse is estimated to have 2ml blood, thus the maximal IV injection volume is 200ul. Use a 1 cc syringe and 26G needle or an insulin syringe with 28G needle. Make sure no airbubbles are present in the syringe. Place the animals under a heating lamp for about 5 minutes or soak the tail in warm water to dilate the tail veins. Put the mouse in a restrainer. Wipe tail with ethanol. Inject the dose slowly
Immunogen: Synthetic peptide corresponding to residues surrounding amino acid 392 of mouse caspase-8
Accession#: BC006737
Gene ID: 12370
Appearance: Colorless liquid
Concentration: 0.5 mg/ml
Formulation: 100 µg (0.5 mg/ml) affinity purified rabbit polyclonal antibody in PBS containing 50% glycerol, 0.5% BSA, and 0.01% thimerosal.
Purification: Affinity purified
Species Reactivity: mouse, rat
Application: Western blot
Application & Usage: Western blotting (0.5-4 µg/ml) in samples from mouse and rat origins. However, the optimal conditions should be determined individually. The antibody detects the 25 kDa large subunit of caspase-8. It does not detect the full-length of caspase-8.
Storage Temp.: -20 °C
Shipping: gel pack
Background Descriptions: Caspase-8, a member of the caspase-family of proteases, plays a key role in mediating Fas (CD95) and TNF induced apoptosis. Caspase-8 is synthesized as inactive pro-enzyme and activation of the enzyme involves proteolytic cleavage that leads to the release of the active p18 and p10 subunits. Activated caspase-8 is able to cleave and activate downstream caspases, such as caspase-3, -6, -7 and a death agonist member of the Bcl-2/Bcl-xL family, Bid, leading to apoptosis.
Handling: The antibody solution should be gently mixed before use.
Usage: For Research Use Only! Not to be used in humans.
Immunogen: Synthetic peptide corresponding to residues surrounding amino acid 392 of mouse caspase-8
Accession#: BC006737
Gene ID: 12370
Appearance: Colorless liquid
Concentration: 0.5 mg/ml
Formulation: 100 µg (0.5 mg/ml) affinity purified rabbit polyclonal antibody in PBS containing 50% glycerol, 0.5% BSA, and 0.01% thimerosal.
Purification: Affinity purified
Species Reactivity: mouse, rat
Application: Western blot
Application & Usage: Western blotting (0.5-4 µg/ml) in samples from mouse and rat origins. However, the optimal conditions should be determined individually. The antibody detects the 25 kDa large subunit of caspase-8. It does not detect the full-length of caspase-8.
Storage Temp.: -20 °C
Shipping: gel pack
Background Descriptions: Caspase-8, a member of the caspase-family of proteases, plays a key role in mediating Fas (CD95) and TNF induced apoptosis. Caspase-8 is synthesized as inactive pro-enzyme and activation of the enzyme involves proteolytic cleavage that leads to the release of the active p18 and p10 subunits. Activated caspase-8 is able to cleave and activate downstream caspases, such as caspase-3, -6, -7 and a death agonist member of the Bcl-2/Bcl-xL family, Bid, leading to apoptosis.
Handling: The antibody solution should be gently mixed before use.
Usage: For Research Use Only! Not to be used in humans.
Apoptosis, Cell Death, and Cell Proliferation Many assays exist to measure apoptosis, cell death and cell proliferation. However, if you have only recently become interested in this area, you may find the diversity of assays for measuring apoptosis, cell death, and cell proliferation bewildering. Here we present our Apoptosis Assay Selection Guide to help you choose the best assays for your purposes.
Kit Summary: • Detection method- Luminometer or Beta Counter. • Sample type- Cell and Tissue lysates, culture media, urine, soil, sludge, plasma and serum, as well as many other biological fluids • Species reactivity- Mammalian • Kit size- 200, 1000 assays • Applications- Bioluminescent detection of the ATP level via luciferase catalyzed reaction for a rapid screening of apoptosis and cell viability in mammalian cells.
Features & Benefits: • Simple one-step procedure; takes only 30 minutes • Fast and convenient • The assay can be done directly in culture plates requiring no harvest/washing/or sample preparations. The assay can be fully automatic for high throughput (10 seconds/sample) and is highly sensitive (detects 10-100 mammalian cells/well).
Kit components: • Nucleotide Releasing Buffer • ATP Monitoring Enzyme • Enzyme Reconstitution Buffer • ATP (MW 551)
Description: Cell death (especially apoptosis) is an energy-dependent process that requires ATP. As ATP levels fall to a point where the cell can no longer perform basic metabolic functions, the cell will die. A typical apoptotic cell exhibits a significant decrease in ATP level. Therefore, loss of ATP level in cell has been used as an indicator of cell death. In contrast, cell proliferation has been recognized by increased levels of ATP. The ApoSENSOR™ Cell Viability Assay Kit utilizes bioluminescent detection of the ATP levels for a rapid screening of apoptosis and cell proliferation simultaneously in mammalian cells. The assay utilizes luciferase to catalyze the formation of light from ATP and luciferin, and the light can be measured using a luminometer or Beta Counter. The assay can be fully automatic for high throughput (10 seconds/sample) and is extremely sensitive (detects 10-100 mammalian cells/well). The high sensitivity of this assay has led to many other applications for detecting ATP production in various enzymatic reactions, as well as for detecting low level bacterial contamination in samples such as blood, milk, urine, soil, and sludge.
Storage Conditions: -70°C
Shipping Conditions: gel pack
USAGE: For Research Use Only! Not For Use in Humans.
Flow cytometry analysis of viable cells. Product can be used in conjunction with R-PE (R-phycoerythrin) and FITC (fluorescein) antibodies in 2-color analysis due to minimal emissions overlap. Product can also be used as a vital dye with the following SouthernBiotech Annexin V conjugates:
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