Revision 8

#64740Store at +4C

1 Kit

(96 assays)

Species Cross Reactivity

H

UniProt ID:

#P22607

Entrez-Gene Id:

#2261

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Color Storage Temp
FGFR3 Rabbit mAb Coated Microwells 67593 96 tests +4C
Phospho Tyrosine Mouse Detection mAb 12982 1 ea Green (Lyophilized) +4C
Anti-mouse IgG, HRP-linked Antibody (ELISA Formulated) 13304 1 ea Red (Lyophilized) +4C
Detection Antibody Diluent 13339 5.5 ml Green +4C
HRP Diluent 13515 5.5 ml Red +4C
Luminol/Enhancer Solution 84850 3 ml RT
Stable Peroxide Buffer 42552 3 ml RT
Sealing Tape 54503 2 ea +4C
ELISA Wash Buffer (20X) 9801 25 ml +4C
ELISA Sample Diluent 11083 25 ml Blue +4C
Cell Lysis Buffer (10X) 9803 15 ml -20C

*The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.

Description

The PathScan® Phospho-FGF Receptor 3 (panTyr) Chemiluminescent Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of tyrosine-phosphorylated FGFR3 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. An FGFR3 rabbit mAb has been coated on the microwells. After incubation with cell lysates, both phospho- and nonphospho-FGFR3 proteins are captured by the coated antibody. Following extensive washing, a phospho tyrosine mouse mAb is added to detect the captured tyrosine-phosphorylated FGFR3 protein. Anti-mouse 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 tyrosine-phosphorylated FGFR3 protein.

*Antibodies in this kit are custom formulations specific to kit.

Specificity/Sensitivity

PathScan® Phospho-FGF Receptor 3 (panTyr) Chemiluminescent Sandwich ELISA Kit #64740 detects endogenous levels of tyrosine-phosphorylated FGFR3 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.

Background

Fibroblast growth factors (FGFs) produce mitogenic and angiogenic effects in target cells by signaling through cell surface receptor tyrosine kinases. There are four members of the FGF receptor family: FGFR1 (flg), FGFR2 (bek, KGFR), FGFR3, and FGFR4. Each receptor contains an extracellular ligand-binding domain, a transmembrane domain, and a cytoplasmic kinase domain (1). Following ligand binding and dimerization, the receptors are phosphorylated at specific tyrosine residues (2). Seven tyrosine residues in the cytoplasmic tail of FGFR1 can be phosphorylated: Tyr463, 583, 585, 653, 654, 730, and 766. Tyr653 and Tyr654 are important for catalytic activity of activated FGFR and are essential for signaling (3). The other phosphorylated tyrosine residues may provide docking sites for downstream signaling components, such as Crk and PLCγ (4,5).

Activating mutations within fibroblast growth factor receptor 3 (FGFR3) are responsible for human skeletal dysplasias including achondroplasia and the neonatal lethal syndromes thanatophoric dysplasia types I and II (6,7). Several of these same FGFR3 mutations as well as overexpression of FGFR3 proteins have also been identified somatically in human cancers, including multiple myeloma, bladder carcinoma and cervical cancer (8). Thus, FGFR3 may represent a potential target for therapy.

  1. Powers, C.J. et al. (2000) Endocr Relat Cancer 7, 165-97.
  2. Reilly, J.F. et al. (2000) J Biol Chem 275, 7771-8.
  3. Mohammadi, M. et al. (1996) Mol Cell Biol 16, 977-89.
  4. Mohammadi, M. et al. (1991) Mol Cell Biol 11, 5068-78.
  5. Larsson, H. et al. (1999) J Biol Chem 274, 25726-34.
  6. Wilkie, A.O. et al. (2002) Am J Med Genet 112, 266-78.
  7. Yamashita, A. et al. (2014) Nature 513, 507-11.
  8. Miyake, M. et al. (2007) Biochem Biophys Res Commun 362, 865-71.

Background References

    Cross-Reactivity Key

    H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    PathScan is a registered trademark of Cell Signaling Technology, Inc.
    U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    Limited Uses

    Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

    Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

    Revision 8
    #64740

    PathScan® Phospho-FGF Receptor 3 (panTyr) Chemiluminescent Sandwich ELISA Kit

    PathScan® Phospho-FGF Receptor 3 (panTyr) Chemiluminescent Sandwich ELISA Kit: Image 1 Expand Image
    Figure 1. Constitutive tyrosine-phosphorylation of FGFR3 in KMS-11 cells lysed in the presence of phosphatase inhibitors (+) is detected by PathScan® Phospho-FGF Receptor 3 (panTyr) Chemiluminescent Sandwich ELISA Kit #64740. In contrast, a low level of phospho-FGFR3 protein is detected in KMS-11 cells lysed in the absence of phosphatase inhibitors* (-). For the FGFR3-negative RPMI 8226 cells, there is no phospho-FGFR3 detected by this ELISA kit in either presence or absence of phosphatase inhibitors. Immediate light generation with chemiluminescent substrate is shown. *Phosphatase inhibitors include sodium pyrophosphate, β-glycerophosphate, and sodium orthovanadate.
    PathScan® Phospho-FGF Receptor 3 (panTyr) Chemiluminescent Sandwich ELISA Kit: Image 2 Expand Image

    Figure 2. The relationship between protein concentration of phospho or nonphospho lysates and immediate light generation with chemiluminescent substrate as detected by the PathScan® Phospho-FGF Receptor 3 (panTyr) Chemiluminescent Sandwich ELISA Kit is shown. Unstarved KMS-11 cells were cultured (0.8 x 106 cells/ml) and lysed with or without addition of phosphatase inhibitors to the lysis buffer (phospho or nonphospho lysate, respectively).