Anti-Interferon beta antibody (ab10740)

Application notes

ELISA: Use at a concentration of 0.5 - 1 µg/ml, this concentration is determined to detect a limit of 1.2 ng/well of recombinant human IFN beta.
Neut: Use at a concentration of 0.05 - 0.2 µg/ml.
WB: Use at a concentration of 0.1 - 0.2 µg/ml, this concentration is determined using human IFN beta at 5 ng/lane under non-reducing and reducing conditions. Predicted molecular weight: 22.4 kDa.

To measure the ability of the antibody to neutralize the bioactivity of human IFN beta, recombinant human IFN beta was incubated with various concentrations of the antibody and added to confluent cultures of HeLa cells. The assay mixture in a total volume of 100 µl per well, containing antibody at concentrations of 0.01 µg/ml- 100 µg/ml, recombinant human IFN beta at 10 ng/ml was incubated at 37°C for 20-24 hours in a humidified CO₂ incubator. Media was aspirated and an appropriate amount of EMCV was added to each well and incubated at for another 20-24 hours. Cells were then fixed, stained and scored for cytopathic effect by measurement of optical densities in a microplate reader at 540nm.

The ND50 is the concentration of antibody required to yield one-half maximal inhibition of the cytokine activity on a responsive cell line, when the cytokine is present at a concentration just high enough to elicit a maximum response.

Not tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.