Cat. # | Size | Price | Inventory |
---|---|---|---|
7296C | 1 Kit (96 assays) |
When ordering five or more kits, please contact us for processing time and pricing.
Looking for this ELISA kit in a 384-well format? Inquire for availability, processing time, and pricing.
REACTIVITY | H |
Product Includes | Volume | Solution Color | |||
---|---|---|---|---|---|
Phospho-PDGFR alpha (Tyr849) Rabbit mAb Coated Microwells | 96 tests | ||||
PDGF Receptor α Rabbit Detection mAb | 1 ea | Red (Lyophilized) | |||
HRP Diluent | 5.5 ml | Red | |||
TMB Substrate 7004 | 11 ml | ||||
STOP Solution 7002 | 11 ml | ||||
Sealing Tape | 2 ea | ||||
ELISA Wash Buffer (20X) 9801 | 25 ml | ||||
Cell Lysis Buffer (10X) 9803 | 15 ml |
Product Information
The rapid protocol (RP) PathScan® RP Phospho-PDGF Receptor alpha (Tyr849) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of PDGF receptor alpha when phosphorylated at Tyr849 in a reduced assay time of 1.5 hours. Incubation of cell lysate and detection antibody on the coated microwell plate forms a sandwich with PDGF Receptor alpha protein phosphorylated at Tyr849 in a single step. The plate is then extensively washed and TMB reagent is added for signal development. The magnitude of absorbance for the developed color is proportional to the quantity of PDGF Receptor alpha protein phosphorylated at Tyr849. Learn more about all of your ELISA kit options here.
*Antibodies in this kit are custom formulations specific to the kit.
NOTE: This protocol is for PathScan® kits that use an HRP directly conjugated to the detection antibody (Rapid Protocol), rather than a 2-step method where the detection antibody and a secondary-HRP are added sequentially.
NOTE: Prepare solutions with deionized/purified water or equivalent.
For adherent cells
For suspension cells
NOTE: Equilibrate all materials and prepared reagents to room temperature prior to running the assay.
NOTE: Initial color of positive reaction is blue, which changes to yellow upon addition of STOP Solution.
created July 2020
Protocol Id: 2144
Platelet derived growth factor (PDGF) family proteins exist as several disulphide-bonded, dimeric isoforms (PDGF AA, PDGF AB, PDGF BB, PDGF CC, and PDGF DD) that bind in a specific pattern to two closely related receptor tyrosine kinases, PDGF receptor α (PDGFRα) and PDGF receptor β (PDGFRβ). PDGFRα and PDGFRβ share 75% to 85% sequence homology between their two intracellular kinase domains, while the kinase insert and carboxy-terminal tail regions display a lower level (27% to 28%) of homology (1). PDGFRα homodimers bind all PDGF isoforms except those containing PDGF D. PDGFRβ homodimers bind PDGF BB and DD isoforms, as well as the PDGF AB heterodimer. The heteromeric PDGF receptor α/β binds PDGF B, C, and D homodimers, as well as the PDGF AB heterodimer (2). PDGFRα and PDGFRβ can each form heterodimers with EGFR, which is also activated by PDGF (3). Various cells differ in the total number of receptors present and in the receptor subunit composition, which may account for responsive differences among cell types to PDGF binding (4). Ligand binding induces receptor dimerization and autophosphorylation, followed by binding and activation of cytoplasmic SH2 domain-containing signal transduction molecules, such as GRB2, Src, GAP, PI3 kinase, PLCγ, and NCK. A number of different signaling pathways are initiated by activated PDGF receptors and lead to control of cell growth, actin reorganization, migration, and differentiation (5). Tyr751 in the kinase-insert region of PDGFRβ is the docking site for PI3 kinase (6). Phosphorylated pentapeptides derived from Tyr751 of PDGFRβ (pTyr751-Val-Pro-Met-Leu) inhibit the association of the carboxy-terminal SH2 domain of the p85 subunit of PI3 kinase with PDGFRβ (7). Tyr740 is also required for PDGFRβ-mediated PI3 kinase activation (8).
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