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Recombinant full length protein (Human).
Our Abpromise guarantee covers the use of ab10516 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||Use a concentration of 8 - 25 µg/ml.|
|Neutralising||Use at an assay dependent concentration.
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 ty
|Inhibition Assay||Use a concentration of 0.02 - 0.08 µg/ml.
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.
|ELISA||Use a concentration of 0.5 - 1 µg/ml.
Concentration determined using 25ng/well recombinant human TRAIL.
|WB||Use a concentration of 1 - 2 µg/ml. Detects a band of approximately 24 kDa (predicted molecular weight: 33.7 kDa). PubMed: 21419121
Determined using human TRAIL at approximately 50 ng/lane under non-reducing and reducing conditions.