Anti-Kit [79D]

Absolute Antibody

Anti-Kit [79D]

7

The crystal structure of the Fab in complex with KIT D4D5 fragment was determined. Surface plasmon resonance analysis was employed to quantitatively characterize the kinetics and dissociation constants of the Fab (Kd = 6.4 pM). 3T3 cells expressing WT KIT receptor were incubated with the IgG and Fab versions of the antibody following stimulation with SCF. Lysates of unstimulated or SCF-stimulated cells were subjected to immunoprecipitation with the IgG and Fab followed by SDS/PAGE and immunoblotting with IgG and Fab. SCF-stimulated autophosphorylation of KIT was strongly inhibited upon binding of both fragments (5 nM for Fab and 1 nM for IgG). The IgG and Fab fragments inhibited the proliferation of KIT-Dependent Ba/F3 cells. The Fab fragment showed inhibition of cell growth at 5 nM, while the IgG fragment at 10 nM (Reshetnyak et al. 2013; PMID: 24127596).

ImmunoPrecipitation, Western Blot, Neutralisation/Blocking, Other Application

Crystallography; SPR; inhibition; IP; WB

Research Use Only

Human

Phage pools consisting of a phage-displayed synthetic antibody library (library F) were cycled through five rounds of selections by using KITD4-5.

-20°C,2°C to 8°C

12352203