Recombinant full length protein (Human).
Abcam is committed to meeting high standards of ethical manufacturing and has decided to discontinue this product by June 2019 as it has been generated by the ascites method. We are sorry for any inconvenience this may cause. We would recommend antibody ab9959 as a replacement.
Our Abpromise guarantee covers the use of ab10516 in the following tested applications.
|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.