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Render Timestamp: 2024-11-21T12:57:41.372Z
Commit: 5c4accf06eb7154018ba3f54329c7590f97f534a
XML generation date: 2024-10-17 20:30:07.049
Product last modified at: 2024-09-19T22:13:02.355Z
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PDP - Template Name: ELISA Kit
PDP - Template ID: *******bd382c2

PathScan® Total EGF Receptor Sandwich ELISA Kit #7250

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  • ELISA

    Supporting Data

    REACTIVITY H
    Application Key:
    • ELISA-ELISA 
    Species Cross-Reactivity Key:
    • H-Human 

    Product Information

    Product Description

    CST's PathScan® Total EGF Receptor Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of total EGF Receptor protein. An EGF Receptor Mouse mAb has been coated onto the microwells. After incubation with cell lysates, both phospho- and nonphospho-EGF Receptor proteins are captured by the coated antibody. Following extensive washing, EGF Receptor Rabbit Antibody is added to detect both the captured phospho- and nonphospho-EGF Receptor protein. Anti-rabbit 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 total EGF Receptor protein.

    *Antibodies in kit are custom formulations specific to kit.

    Protocol

    Specificity / Sensitivity

    CST's PathScan® Total EGF Receptor Sandwich ELISA Kit #7250 detects endogenous levels of EGF Receptor Protein. As shown in Figure 1, using PathScan® Phospho-EGF Receptor (Tyr1068) ELISA Kit #7240, a significant induction of Phospho-EGF Receptor (Tyr1068) is detected in A-431 cells treated with EGF. The levels of total EGF Receptor (phospho and non-phospho) detected by PathScan® Total EGF Receptor Sandwich ELISA Kit #7250 remain unchanged. In Figure 3, western blot analysis of protein captured in the EGF Receptor mouse mAb coated microwell shows a single band corresponding to the EGF Receptor. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

    Species Reactivity:

    Human

    Background

    The epidermal growth factor (EGF) receptor is a transmembrane tyrosine kinase that belongs to the HER/ErbB protein family. Ligand binding results in receptor dimerization, autophosphorylation, activation of downstream signaling, internalization, and lysosomal degradation (1,2). Phosphorylation of EGF receptor (EGFR) at Tyr845 in the kinase domain is implicated in stabilizing the activation loop, maintaining the active state enzyme, and providing a binding surface for substrate proteins (3,4). c-Src is involved in phosphorylation of EGFR at Tyr845 (5). The SH2 domain of PLCγ binds at phospho-Tyr992, resulting in activation of PLCγ-mediated downstream signaling (6). Phosphorylation of EGFR at Tyr1045 creates a major docking site for the adaptor protein c-Cbl, leading to receptor ubiquitination and degradation following EGFR activation (7,8). The GRB2 adaptor protein binds activated EGFR at phospho-Tyr1068 (9). A pair of phosphorylated EGFR residues (Tyr1148 and Tyr1173) provide a docking site for the Shc scaffold protein, with both sites involved in MAP kinase signaling activation (2). Phosphorylation of EGFR at specific serine and threonine residues attenuates EGFR kinase activity. EGFR carboxy-terminal residues Ser1046 and Ser1047 are phosphorylated by CaM kinase II; mutation of either of these serines results in upregulated EGFR tyrosine autophosphorylation (10).
    For Research Use Only. Not For Use In Diagnostic Procedures.
    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.
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