Cell Signaling Technology

Product Pathways - Tyrosine Kinase/ Adaptors

Phospho-FLT3 (Tyr591) Antibody (Alexa Fluor® 488 Conjugate) #3459

Applications Reactivity Source
F H Rabbit

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

Specificity / Sensitivity

Phospho-FLT3 (Tyr591) Antibody detects endogenous FLT3 only when phosphorylated at tyrosine 591. The antibody does not cross-react with other FLT family members. It may cross-react with some tyrosine phosphorylated proteins. (Patent Pending.)

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with a synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding Tyr591 of human FLT3. Antibodies are purified by protein A and peptide affinity chromatography.The antibody was conjugated to Alexa Fluor®488 under optimal conditions with an F/P ratio of 2-5.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of SEM cells, treated with a FLT3 inhibitor (blue) or untreated (green), using Phospho-FLT3 (Tyr591) Antibody (Alexa Fluor® 488 Conjugate).

Description

This Cell Signaling Technology Antibody was conjugated to Alexa Fluor®488 fluorescent dye and tested in-house for direct flow cytometric analysis of human cells. The unconjugated antibody #3461 reacts with human and mouse Phospho-FLT3 (Tyr591). CST expects that Phospho-FLT3 (Tyr591) Antibody (Alexa Fluor® 488 Conjugate) will also recognize Phospho-FLT3 (Tyr591) in these species.

Background

FMS-related tyrosine kinase 3 (FLT3, also called Flk2), is a member of the type III receptor tyrosine kinase family, which includes c-Kit, PDGFR and M-CSF receptors. FLT3 is expressed on early hematopoietic progenitor cells and supports growth and differentiation within the hematopoietic system (1,2). FLT3 is activated after binding with its ligand FL, which results in a cascade of tyrosine autophosphorylation and tyrosine phosphorylation of downstream targets (3). The p85 subunit of PI3 kinase, SHP2, GRB2 and Shc are associated with FLT3 after FL stimulation (4-6). Tyr589/591 is located in the juxtamembrane region of FLT3 and may play an important role in regulation of FLT3 tyrosine kinase activity. Somatic mutations of FLT3 consisting of internal tandem duplications (ITDs) occur in 20% of patients with acute myeloid leukemia (7).

  1. Shurin, M.R. et al. (1998) Cytokine Growth Factor Rev. 9, 37-48.
  2. Naoe, T. et al. (2001) Cancer Chemother. Pharmacol. 48 Suppl1, S27-S30.
  3. Namikawa, R. et al. (1996) Stem Cells 14, 388-395.
  4. Beslu, N. et al. (1996) J. Biol. Chem. 271, 20075-20081.
  5. Zhang, S. and Broxmeyer, H.E. (2000) Biochem. Biophys. Res. Commun. 277, 195-199.
  6. Zhang, S. et al. (1999) J. Leukoc. Biol. 65, 372-380.
  7. Mizuki, M. et al. (2000) Blood 96, 3907-3914.

Application References

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