Cat. No.
MABL-2204
Application
crystallography, inhibition, IP, SPR, WB
Isotype
Engineer antibody
Species Reactivity
Human
Clone No.
79D
From
Recombinant Antibody
Specificity
The antibody is specific for KIT, and it binds to the membrane-proximal Ig-like D4 of KIT. The receptor tyrosine kinase KIT is a transmembrane protein that plays crucial roles in mediating diverse cellular processes, including cell differentiation, proliferation, and cell survival, among other activities.
Alternative Names
Mast/stem cell growth factor receptor Kit (EC:2.7.10.1); SCFR; Piebald trait protein; PBT; Proto-oncogene c-Kit; Tyrosine-protein kinase Kit; p145 c-kit; v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene homolog; CD117
UniProt
P10721
Immunogen
Phage pools consisting of a phage-displayed synthetic antibody library (library F) were cycled through five rounds of selections by using KITD4-5.
Application Notes
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).
Antibody First Published
Reshetnyak et al. Structural basis for KIT receptor tyrosine kinase inhibition by antibodies targeting the D4 membrane-proximal region. Proc Natl Acad Sci U S A. 2013 Oct 29; 110(44): 17832–17837. PMID:24127596
Note on publication
The paper describes the generation and characterization of a panel of inhibitory KIT antibodies.
Size
100 μg Purified antibody.
Concentration
1 mg/ml.
Purification
Protein A affinity purified
Buffer
PBS with 0.02% Proclin 300.
Storage Recommendation
Store at 4⁰C for up to 3 months. For longer storage, aliquot and store at - 20⁰C.
