Anti-Macrophage Inflammatory Protein 3 beta antibody (ab10389)

Application notes

ELISA: Use at a concentration of 0.5 - 1 µg/ml to detect a limit of 0.16 ng/well of recombinant mouse MIP3 beta..
IHC-P: Use at an assay dependent dilution (PMID 19917684).
Neut: Use at a concentration of 1 - 3 µg/ml to neutralize 50% of the bioactivity due to 0.1 ug/ml recombinant mouse MIP3 beta using cultured lymphocytes.
WB: Use at a concentration of 0.1 - 0.2 µg/ml using mouse MIP3 beta at 1 ng/lane under non-reducing and reducing conditions. Predicted molecular weight: 11 kDa.

To measure the ability of the antibody to neutralize the bioactivity of mouse MIP3 beta, recombinant mouse MIP3 beta was incubated with various concentrations of the antibody for 30 minutes in a microwell plate. Following preincubation, 35 ul of cytokine-antibody solution (containing antibody at concentrations of 0.01- 200 ug/ml and recombinant mouse MIP3 beta at 0.1 ug/ml) was transferred to the lower compartment of a 96-well chemotaxis chamber. The chamber was assembled using a PVP-free polycarbonate filter and 2 x 10⁶ cells/well were added to the top chamber. After the plate was incubated at 37°C for 3 hours in a humidified CO₂ incubator, the cells that migrated through to the lower chamber were stained using MTT and the optical density was measured. 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 yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.