The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
应用说明ELISA: Use at a concentration of 0.5 - 1 µg/ml, determined using 25ng/well recombinant human TRAIL.
WB: Use at a concentration of 1 - 2 µg/ml. Detects a band of approximately 24 kDa (predicted molecular weight: 33.7 kDa) - Determined using human TRAIL at approximately 50 ng/lane under non-reducing and reducing conditions.
Inhib: Use at a concentration of 0.02 - 0.08 µg/ml.
Inhib: Monoclonal anti-TRAIL neutralizes bioactivity on mouse L929 cells.
To measure the ability of this antibody to neutralize the biological activity of TRAIL, cross-linked recombinant human TRAIL is incubated with various concentrations of the antibody for 1 hour at 37°C in a 96 well plate. Following this preincubation period, mouse L929 cells are added. The assay mixture in a total volume of 150 µl per well, containing antibody at concentrations of 0.01-100 ng/ml, cross-linked recombinant human TRAIL at 12 ng/ml and actinomycin D at 1 µg/ml, are incubated at 37°C for 24 hours in a humidified CO2 incubator. Following incubation, the cells are fixed with 5% formaldehyde and then stained with a crystal violet solution containing 100 µl of 33% acetic acid. Absorbance at 540 nm is read on a microplate reader. The exact concentration of antibody required to neutralize TRAIL is dependent on the cytokine concentration, cell type, growth conditions and the type of activity.
The Neutralization Dose50 (ND50) for this antibody is defined as that concentration required to yield one-half maximal inhibition of the TRAIL-mediated activity on a responsive cell line, when TRAIL is present at a concentration high enough to elicit a maximum response. Monoclonal anti-TRAIL (APO-2L) has the ability to neutralize the biological activity of recombinant human TRAIL. The ND50 for this effect is approximately 2-8 ng/ml in the presence of 12 ng/ml of cross-linked recombinant human TRAIL, using mouse L929 cells.
Not tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
功能Cytokine that binds to TNFRSF10A/TRAILR1, TNFRSF10B/TRAILR2, TNFRSF10C/TRAILR3, TNFRSF10D/TRAILR4 and possibly also to TNFRSF11B/OPG. Induces apoptosis. Its activity may be modulated by binding to the decoy receptors TNFRSF10C/TRAILR3, TNFRSF10D/TRAILR4 and TNFRSF11B/OPG that cannot induce apoptosis.
组织特异性Widespread; most predominant in spleen, lung and prostate.
Tumor necrosis factor ligand superfamily member 10 antibody
Anti-TRAIL antibody [75411.11] (ab10516)参考文献
This product has been referenced in:
Hicks SD & Miller MW Effects of ethanol on transforming growth factor ?1-dependent and -independent mechanisms of neural stem cell apoptosis. Exp Neurol229:372-80 (2011).
Read more (PubMed: 21419121) »
Roux S et al. CD4+CD25+ Tregs control the TRAIL-dependent cytotoxicity of tumor-infiltrating DCs in rodent models of colon cancer. J Clin Invest118:3751-61 (2008).
Read more (PubMed: 18830416) »