Ordering Information

Catalogue Number : 17-490
Qty/Pk : 96 assays

ErbB2 (Tyr1248) Dual Detect CELISA Assay Kit (Fluorogenic Detection)

Description:

Trade Name:

Dual Detect CELISA

Qty/Pk:

96 assays

Product Overview:

Dual Detect CELISA is a straightforward and efficient cell-based ELISA assay that eliminates the need to prepare cell lysates. It allows for the measurement of the relative expression levels of both the total and the phosphorylated forms of the target protein in whole cells in a convenient 96 well microplate format using dual fluorescent detection. This allows for the normalization of total protein between wells, thus allowing a more accurate measurement of phosphorylation levels among cell and/or growth condition differentials.
Cells are cultured directly in the microplate and stimulated as desired. They are then fixed and simultaneously incubated with two primary antibodies. One is a phosphospecific antibody directed against the phosphorylated epitope of the protein that correlates with it activation and the other is a total antibody that recognizes the target-protein, regardless of phosphorylation status. The levels of both the total target protein and the phosphorylated protein are measured simultaneously in a single well using a double immunoenzymatic labeling procedure, either horseradish-peroxidase (HRP) or alkaline phosphatase (AP), and two spectrally distinct fluorogenic substrates for HRP and AP. The fluorescence of the phosphorylated protein is normalized to the total protein in each well thus reducing well-to-well variations. The ErbB2 (Try1248) kit contains the components required to run the Dual Detect CELISA using fluorogenic substrates to measure both total ErbB2 and phosphorylated ErbB2 (Tyr1248) in the context of whole cells.

Background Information:

ErbB2, also known as HER2/Neu, is a key receptor tyrosine kinase that is one of the four members of the EGFR family of kinases. v-ErbB was identified as the viral counterpart of the EGF Receptor, and its relatives have adopted this nomenclature (or the HER nomenclature for human proteins). These proteins are activated by various growth factors such as Heregulin, Neuregulin, and Amphiregulin. All the family members are capable of homo- and heterodimerization, and together activate various downstream signaling pathways. erbB-2/HER-2/neu is amplified in highly aggressive breast cancers and has become the target of multiple drugs. All members of this family, when Tyrosine phosphorylated, can nucleate the formation of signaling complexes through adapter proteins.

Species Reactivity: