Product Pathways - Tyrosine Kinase / Adaptors
PathScan® Phospho-FGF Receptor 4 (panTyr) Sandwich ELISA Kit #69193
|Product Includes||Volume||Solution Color|
|ELISA Sample Diluent||25 ml||Blue|
|Phospho Tyrosine Mouse Detection mAb||1 ea||Green (Lyophilized)|
|Anti-mouse IgG, HRP-linked Antibody (ELISA Formulated)||1 ea||Red (Lyophilized)|
|Detection Antibody Diluent||11 ml||Green|
|HRP Diluent||11 ml||Red|
|STOP Solution||11 ml||Colorless|
|TMB Substrate #7004||11 ml||Colorless|
|ELISA Wash Buffer (20X)||25 ml||Colorless|
|Cell Lysis Buffer (10X) #9803||15 ml||Yellowish|
|FGFR4 Rabbit mAb Coated Microwells||1 ea|
|Sealing Tape||2 sheets|
Specificity / Sensitivity
PathScan® Phospho-FGF Receptor 4 (panTyr) Sandwich ELISA Kit recognizes endogenous levels of tyrosine-phosphorylated FGF receptor 4 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.
PathScan® Phospho-FGF Receptor 4 (panTyr) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of tyrosine-phosphorylated FGR receptor 4 protein. An FGF receptor 4 rabbit mAb has been coated onto the microwells. After incubation with cell lysates, FGF receptor 4 protein (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, a phospho-tyrosine mouse detection mAb is added to detect captured phospho-FGF receptor 4 protein. Anti-mouse IgG, HRP-linked antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of optical density for this developed color is proportional to the quantity of FGF Receptor 4 protein phosphorylated on tyrosine residues.
Antibodies in the kit are custom formulations specific to kit.
Figure 2: The relationship between protein concentration of lysates from untreated and hFGF-treated Hep G2 cells and the absorbance at 450 nm as detected by the PathScan® Phospho-FGF Receptor 4 (panTyr) Sandwich ELISA Kit #69193 is shown. Starved Hep G2 cells (85% confluence) were treated with Human FGF acidic (hFGF acidic) #5234 (100 ng/ml, 2-5 min, 37°C) and then lysed.
ELISA - Western correlation
Figure 1: Treatment of Hep G2 cells with Human FGF acidic (hFGF acidic) #5234 stimulates tyrosine phosphorylation of FGF receptor 4 protein, which can be detected by the PathScan® Phospho-FGF Receptor 4 (panTyr) Sandwich ELISA Kit. This kit can also detect the high level of tyrosine-phosphorylated FGF receptor 4 from CHO cells that are transfected with human FGF receptor 4 protein. The absorbance readings at 450 nm are shown in the top figure while corresponding western blots using FGF Receptor 4 (D3B12) XP® Rabbit mAb #8562 (left) and Phospho-FGF Receptor (Tyr653/654) (55H2) Mouse mAb #3476 (right) are shown in the bottom figure.
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).
- Powers, C.J. et al. (2000) Endocr Relat Cancer 7, 165-97.
- Reilly, J.F. et al. (2000) J Biol Chem 275, 7771-8.
- Mohammadi, M. et al. (1996) Mol Cell Biol 16, 977-89.
- Mohammadi, M. et al. (1991) Mol Cell Biol 11, 5068-78.
- Larsson, H. et al. (1999) J Biol Chem 274, 25726-34.
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- 15078 PathScan® Total FGF Receptor 4 Sandwich ELISA Kit
- 3476 Phospho-FGF Receptor (Tyr653/654) (55H2) Mouse mAb
- 7004 TMB Substrate
- 7076 Anti-mouse IgG, HRP-linked Antibody
- 8562 FGF Receptor 4 (D3B12) XP® Rabbit mAb
- 9803 Cell Lysis Buffer (10X)
- 9808 Phosphate Buffered Saline (PBS-20X)
- 9809 Phosphate Buffered Saline with Tween® 20 (PBST-20X)
- 9998 BSA
For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
XP is a registered 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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