Anti-Lymphotactin antibody (ab10607)

Application notes

ELISA: Use at a concentration of 0.5 - 1 µg/ml. The detection limit for recombinant human lymphotactin is approximately 0.3 ng/well.
Neut: Use at a concentration of 20 - 30 µg/ml.
WB: Use at a concentration of 1 - 2 µg/ml. The detection limit for recombinant human lymphotactin is approximately 5 ng/lane under non-reducing and reducing conditions.Predicted molecular weight: 13.6 kDa.

To measure the ability of anti-human lymphotactin to neutralize the bioactivity of recombinant human lymphotactin for BaF/3 human XCR-1 transfected cells, recombinant human lymphotactin is incubated with various concentrations of the antibody for 15 minutes at room temperature in a 96 well plate. Following this pre-incubation, 75 µl of the cytokine-antibody solution (containing recombinant human lymphotactin at a final concentration of 0.5 µg/ml and antibody at concentrations from 0.1-200 µg/ml) is transferred to the lower compartment of a 96 well chemotaxis chamber. The chemotaxis chamber is then assembled using a PVP-free polycarbonate filter (5 micron size) and 2 x 10⁶ cells/well is added to the top chamber. Following incubation for 3 hours at 37 °C in a 5% CO₂ humidified incubator, the chamber is carefully disassembled. The cells that migrate through to the lower chamber are transferred to a 96 well plate. Chemotaxis is measured by Alamar Blue staining of cells that have migrated through the filter.

The Neutralization Dose50 (ND50) for anti-human lymphotactin is approximately 20 to 30 µg/ml in the presence of 0.5 µg/ml of recombinant human lymphotactin using BaF/3 cells transfected with recombinant human XCR-1. 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. The exact concentration of antibody required to neutralize recombinant human lymphotactin activity is dependent on the cytokine concentration, cell type, growth conditions, and the type of activity studied.

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