Cell Signaling Technology

Product Pathways - Tyrosine Kinase / Adaptors

PathScan® Total FGF Receptor 3 Sandwich ELISA Kit #15077

fgf   FGF receptor 3   fgfr   fgfr-3   fgfr3   flg   sandwich elisa   screen  

REACTIVITY
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No. Size Price
15077C 1 Kit ( 96 assays ) ¥6,345.00 现货查询 购买询价
Product Includes Volume Solution Color
ELISA Sample Diluent 25 ml Blue
Anti-rabbit IgG, HRP-linked Antibody (ELISA Formulated) 1 ea Red (Lyophilized)
Detection Antibody Diluent 11 ml Green
HRP Diluent 11 ml Red
FGF Receptor 3 Rabbit Detection mAb 1 ea Green (Lyophilized)
FGFR3 Mouse mAb Coated Microwells 1 ea
STOP Solution #7002 11 ml Colorless
TMB Substrate #7004 11 ml Colorless
ELISA Wash Buffer (20X) 25 ml Colorless
Cell Lysis Buffer (10X) #9803 15 ml Yellowish
Sealing Tape 2 sheets

Specificity / Sensitivity

PathScan® Total FGF Receptor 3 Sandwich ELISA Kit detects endogenous levels of FGFR3 protein in human cells, as shown in Figure 1. The kit sensitivity is shown in Figure 2. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Description

The PathScan® Total FGF Receptor 3 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of FGF receptor 3 (FGFR3) protein. An FGFR3 mouse antibody has been coated onto the microwells. After incubation with cell lysates, both phospho- and nonphospho-FGFR3 proteins are captured by the coated antibody. Following extensive washing, an FGFR3 rabbit antibody is added to detect captured FGFR3 proteins. An anti-rabbit IgG, HRP-linked antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added for colorimetric detection. The magnitude of absorbance for the developed color is proportional to the quantity of FGFR3 protein.

Antibodies in the kit are custom formulations specific to the kit.

ELISA - Western correlation

ELISA - Western correlation

Figure 1. FGF receptor 3 protein is detected at varying levels from multiple cell lines using the PathScan® Total FGF Receptor 3 Sandwich ELISA Kit #15077. The absorbance readings at 450 nm are shown in the top figure, while corresponding western blots using FGF Receptor 3 (C51F2) Rabbit mAb #4574 are shown in the bottom figure.

Sensitivity

Sensitivity

Figure 2. The relationship between protein concentration of lysates from KMS-11 cells and the absorbance at 450 nm as detected by the PathScan® Total FGF Receptor 3 Sandwich ELISA Kit #15077 is shown. KMS-11 cells (0.5-1.0 x 106 cells/ml) were harvested and then lysed.

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 (6,8). 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 (7). 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. Miyake, M. et al. (2007) Biochem Biophys Res Commun 362, 865-71.
  8. Yamashita, A. et al. (2014) Nature 513, 507-11.

Application References

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Protocols

Companion Products


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

U.S. Patent No. 5,675,063.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

PathScan is a trademark of Cell Signaling Technology, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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