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28435
FLT3 (BV10A4H2) Mouse mAb (Pacific Blue™ Conjugate)
Antibody Conjugates

FLT3 (BV10A4H2) Mouse mAb (Pacific Blue™ Conjugate) #28435

APPLICATIONS

REACTIVITY SENSITIVITY MW (kDa) Isotype
H Endogenous Mouse IgG1
Flow Cytometry

Flow cytometric analysis of live K-562 cells (blue) and SEM cells (green) using FLT3 (BV10A4H2) Mouse mAb (Pacific Blue™ Conjugate) (solid lines) or concentration-matched conjugated Mouse (G3A1) mAb IgG1 Isotype Control (Pacific Blue™ Conjugate) #14411 (dashed lines).

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Flow Cytometry

A. Solutions and Reagents

NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.

  1. 20X Phosphate Buffered Saline (PBS): (#9808) To prepare 1 L 1X PBS: add 50 ml 20X PBS to 950 ml dH2O, mix.
  2. Incubation Buffer: Dissolve 0.5 g Bovine Serum Albumin (BSA) (#9998) in 100 ml 1X PBS. Store at 4°C.

B. Immunostaining

  1. Count cells using a hemacytometer or alternative method.
  2. Aliquot 0.5-1x106 cells into each assay tube (by volume).
  3. Add 2-3 ml Incubation Buffer to each tube and rinse by centrifugation. Repeat.
  4. Resuspend cells in 100 µl of diluted primary antibody (prepared in incubation buffer at the recommended dilution).
  5. Incubate for 1 hr at room temperature.
  6. Wash by centrifugation in 2-3 ml incubation buffer.
  7. Resuspend cells in the appropriate volume of PBS and analyze on flow cytometer.

posted June 2017

Protocol Id: 1504

Application Dilutions
Flow Cytometry 1:50
Storage:

Supplied in PBS (pH 7.2), less than 0.1% sodium azide and 2 mg/ml BSA. Store at 4°C. Do not aliquot the antibody. Protect from light. Do not freeze.

FLT3 (BV10A4H2) Mouse mAb (Pacific Blue™ Conjugate) recognizes endogenous levels of total FLT3 protein.

Species Reactivity:

Human

BV10A4H2 clone is produced by immunizing animals with the leukemic cell line BV-173 and Antibody is purified from tissue culture supernatant via protein G chromatography.

This Cell Signaling Technology antibody is conjugated to Pacific Blue™ fluorescent dye and tested in-house for direct flow cytometric analysis in human cells.

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 Suppl 1, S27-30.
  3. Namikawa, R. et al. (1996) Stem Cells 14, 388-95.
  4. Beslu, N. et al. (1996) J Biol Chem 271, 20075-81.
  5. Zhang, S. and Broxmeyer, H.E. (2000) Biochem Biophys Res Commun 277, 195-9.
  6. Zhang, S. et al. (1999) J Leukoc Biol 65, 372-80.
  7. Mizuki, M. et al. (2000) Blood 96, 3907-14.
Entrez-Gene Id
2322
Swiss-Prot Acc.
P36888
For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Pacific Blue is a trademark of Molecular Probes, Inc.

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To Purchase # 28435S

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Product # Size Price
28435S
100 µl  (50 tests) $ 305