The AlphaLISA® Human epidermal growth factor receptor 2 (ErbB-2, NEU, or HER2) Detection Kit is designed for detection and quantitation of human ErbB-2 in serum, buffered solution or cell culture medium in a homogeneous (no-wash steps, no separation steps) assay.
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The antibodies target the extracellular domain of the HER2 receptor. The analyte in this kit consists of the extracellular domain of human HER2 fused to the Fc region of human IgG1.
Human epidermal growth factor receptor 2 (ERBB2, NEU, or HER2) is a type I membrane glycoprotein that is a member of the ERBB family of tyrosine kinase receptors. It serves as a receptor for the epidermal growth factor (EGF) family of growth factors. HER2 is widely expressed in epithelial cells and has also been found to be over-expressed in a large number of breast carcinomas. It is an orphan receptor with no known ligand, as it acts as a heterodimer with other members of the EGF receptors family. HER2 appears to play roles in development, cancer, communication at the neuromuscular junction, and regulation of cell growth and differentiation. HER2 can be shed from the cell surface as a soluble protein (sHER2) via proteolytic cleavage by an unidentified protease. sHER2 seems to be an important serum biomarker in certain types of cancer.
AlphaLISA technology allows the detection of molecules of interest in a no-wash, highly sensitive, quantitative assay. In an AlphaLISA assay, a biotinylated anti-analyte antibody binds to the Streptavidin-coated Donor beads while another anti-analyte antibody is conjugated to AlphaLISA Acceptor beads. In the presence of the analyte, the beads come into close proximity. The excitation of the Donor beads causes the release of singlet oxygen molecules that triggers a cascade of energy transfer in the Acceptor beads, resulting in a sharp peak of light emission at 615 nm.
|Assay Target Class||Protein|
|Experimental Type||In vitro|
|Product Brand Name||AlphaLISA|
|Shipping Condition||Blue Ice|
|Unit Size||5,000 assay points|
Biomarker levels were measured directly in CulturPlates-96 and-384 in a simple, fast, all-in-one-well AlphaLISA assay format. The elimination of transfer and wash steps simplifies cellular assays, reduces variability and significantly reduces hands-on time and costs associated with consumables. Integral plasma membrane (EGFR) and secreted solubleproteins (TNFa, IL1ß, IL6, IL8) were successfully determined, on adherent or suspension cells, using the standard AlphaLISA Immunoassay buffer.
The AlphaLISA® assay is a homogeneous immunoassay alternative to classical ELISA. AlphaLISA assays were originally utilized to detect analytesin cell cultures upernatants or serum/plasma samples.
Enzyme-linked Immunosorbent Assay is the most widely Kits adopted method for detection and quantification of cytokines and other biomarkers. This traditional technology offers good,selectivity, sensitivity and assay versatility; however, it has certain disadvantages such as limited dynamic range and low throughput due to the numerous wash steps.