Cell Signaling Technology

Product Pathways - Tyrosine Kinase / Adaptors

c-Kit (Ab81) Mouse mAb (Alexa Fluor® 488 Conjugate) #3310

Applications Reactivity Sensitivity Isotype
IF-IC F H Endogenous Mouse IgG1

Applications Key:  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

c-Kit (Ab81) Mouse mAb detects endogenous levels of total c-Kit protein. It does not cross-react with other receptor tyrosine kinase family members.

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant proteins containing the cytoplasmic domain of human c-Kit. The antibody was conjugated to Alexa Fluor® 488 under optimal conditions with an F/P ratio of 2-6.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat (blue) and H526 (green) cells using c-Kit (Ab81) Mouse mAb (Alexa Fluor® 488 Conjugate).

IF-IC

IF-IC

Confocal immunofluorescent analysis of H526 cells using c-Kit (Ab81) Mouse mAb (Alexa Fluor® 488 Conjugate) (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Description

This Cell Signaling Technology antibody is conjugated to Alexa Fluor ® 488 fluorescent dye and tested in-house for direct flow cytometry and immunofluorescent analysis in human cells.

Background

c-Kit is a member of the subfamily of receptor tyrosine kinases that includes PDGF, CSF-1, and FLT3/flk-2 receptors (1,2). It plays a critical role in activation and growth in a number of cell types including hematopoietic stem cells, mast cells, melanocytes, and germ cells (3). Upon binding with its stem cell factor (SCF) ligand, c-Kit undergoes dimerization/oligomerization and autophosphorylation. Activation of c-Kit results in the recruitment and tyrosine phosphorylation of downstream SH2-containing signaling components including PLCγ, the p85 subunit of PI3 kinase, SHP2, and CrkL (4). Molecular lesions that impair the kinase activity of c-Kit are associated with a variety of developmental disorders (5), and mutations that constitutively activate c-Kit can lead to pathogenesis of mastocytosis and gastrointestinal stromal tumors (6). Tyr719 is located in the kinase insert region of the catalytic domain. c-Kit phosphorylated at Tyr719 binds to the p85 subunit of PI3 kinase in vitro and in vivo (7).

  1. Martin, F.H. et al. (1990) Cell 63, 203-11.
  2. Yarden, Y. et al. (1987) EMBO J 6, 3341-51.
  3. Gommerman, J.L. et al. (1997) J Biol Chem 272, 30519-25.
  4. Sattler, M. et al. (1997) J Biol Chem 272, 10248-53.
  5. Nocka, K. et al. (1990) EMBO J 9, 1805-13.
  6. Hirota, S. et al. (1998) Science 279, 577-80.
  7. Blume-Jensen, P. et al. (2000) Nat Genet 24, 157-62.

Application References

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Alexa Fluor® is a registered trademark of Molecular Probes, Inc.

For Research Use Only. Not For Use In Diagnostic Procedures.

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