Cell Signaling Technology

Product Pathways - PathScan ELISA

PathScan® Phospho-FLT3 (Tyr591) Chemiluminescent Sandwich ELISA Kit #7021

When ordering five or more kits, please contact us for processing time and pricing at sales@cellsignal.com.

Kit Includes Volume Solution Color
FLT3 Mouse mAb coated microwells 96 tests
Phospho-FLT3 (Tyr591) Rabbit Detection Ab 5.5 ml Green
Anti-rabbit IgG, HRP-Linked Ab 5.5 ml Red
Luminol/Enhancer Solution 3 ml Colorless
Stable Peroxide Buffer 3 ml Colorless
Sealing Tape 2 sheets
ELISA Wash Buffer (20X) 25 ml Colorless
ELISA Sample Diluent 25 ml Blue
Cell Lysis Buffer (10X) #9803 15 ml Yellowish

Note: 12 8-well modules –Each module is designed to break apart for 8 tests.
Note: Kit should be stored at 4°C with the exception of Cell Lysis Buffer, which is stored at –20°C (packaged separately).

Species Cross-Reactivity

H

Reactivity Key:  H=Human
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Description

The PathScan® Phospho-FLT3 (Tyr591) Chemiluminescent Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of phospho-FLT3 (Tyr591) protein with a chemiluminescent readout. Chemiluminescent ELISAs often have a wider dynamic range and higher sensitivity than conventional chromogenic detection. This chemiluminescent ELISA, which is offered in low volume microplates, shows increased signal and sensitivity while using a smaller sample size. A FLT3 Mouse mAb has been coated on the microwells. After incubation with cell lysates, FLT3 (phospho or nonphospho) protein is captured by the coated antibody. Following extensive washing, a phospho-FLT3 (Tyr 591) Rabbit Antibody is added to detect the captured phospho-FLT3 protein. Anti-rabbit IgG, HRP-linked antibody is then used to recognize the bound detection antibody. Chemiluminescent reagent is added for signal development. The magnitude of light emission, measured in relative light units (RLU), is proportional to the quantity of phospho-FLT3 (Tyr591) protein.Antibodies in kit are custom formulations specific to kit.

Specificity / Sensitivity

PathScan® Phospho-FLT3 (Tyr591) Chemiluminescent Sandwich ELISA Kit #7021 detects endogenous levels of phospho-FLT3 (Tyr591) in human cells. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

ELISA

ELISA

Relationship between protein concentration of lysates from phospho or nonphospho SEM cells and immediate light generation with chemiluminescent substrate is shown. SEM cells in exponential growth (106 cells/ml) were lysed with or without the addition of phosphatase inhibitor to the lysis buffer (phospho or nonphospho lysate). The graph inset corresponding to the shaded area shows high sensitivity and a linear response at the low protein concentration range.

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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Protocols

* Product-specific protocol.

Companion Products

US Patent No. 7,183,385 and foreign equivalents assigned to Cell Signaling Technology, Inc.

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

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