Product Pathways - PathScan ELISA
PathScan® Total FGFR2 Sandwich ELISA Kit #7939
When ordering five or more kits, please contact us for processing time and pricing at firstname.lastname@example.org.
|Kit Includes||Volume||Solution Color|
|FGFR2 Rabbit Antibody coated microwells||96 tests|
|FGFR2 Mouse Detection Antibody||11 ml||Green|
|Anti-rabbit IgG, HRP- linked Antibody||11 ml||Red|
|TMB Substrate #7004||11 ml||Colorless|
|STOP Solution #7002||11 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 enclosed in parentheses are predicted to react based on 100% sequence homology.
The PathScan® Total FGFR2 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of FGFR2 protein. An FGFR2 rabbit antibody has been coated on the microwells. After incubation with cell lysates, FGFR2 protein (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, a FGFR2 mouse detection antibody is added to detect captured FGFR2 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 the absorbance for this developed color is proportional to the quantity of total FGFR2 protein.Antibodies in kit are custom formulations specific to kit.
Specificity / Sensitivity
PathScan® Total FGFR2 Sandwich ELISA Kit #7939 detects endogenous levels of FGFR2 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.
ELISA - Western correlation
Figure 1. Constitutive phosphorylation of FGFR2 in KATO III cells lysed in the presence of phosphatase inhibitors* (phospho lysate) is detected by PathScan® Phospho-FGFR2 (panTyr) Sandwich ELISA Kit #7954 (upper, right). In contrast, a low level of phospho-FGFR2 protein is detected in KATO III cells lysed in the absence of phosphatase inhibitors (nonphospho lysate). Similar levels of total FGFR2 protein from both nonphospho and phospho lysates are detected by PathScan® Total FGFR2 Sandwich ELISA Kit #7939 (upper, left). Absorbance at 450 nm is shown in the top figure while corresponding western blots using Phospho-FGF Receptor (Tyr653/654) Antibody #3471 (right) or a total FGFR2 Rabbit mAb (left) are shown in the bottom figure. *Phosphatase inhibitors include sodium pyrophosphate, β-glycerophosphate, and Na3VO4.
Figure 2. The relationship between protein concentration of phospho or nonphospho lysates and the absorbance at 450 nm is shown. KATO III cells were cultured (85% confluence) and lysed with or without the addition of phosphatase inhibitors to the lysis buffer (phospho or nonphospho lysate, respectively).
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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- 7954 PathScan® Phospho-FGFR2 (panTyr) Sandwich ELISA Kit
- 3471 Phospho-FGF Receptor (Tyr653/654) Antibody
- 3476 Phospho-FGF Receptor (Tyr653/654) (55H2) Mouse mAb
- 7076 Anti-mouse IgG, HRP-linked Antibody
- 9803 Cell Lysis Buffer (10X)
- 9809 Phosphate Buffered Saline with Tween 20 (PBST-20X)
- 9998 BSA
- 7004 TMB Substrate
- 7002 STOP Solution
For Research Use Only. Not For Use In Diagnostic Procedures.