Anti-TRAIL antibody [75411.11] (ab10516)

Application notes

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.