AlphaLISA® no-wash immunoassay kit for detection of human C-peptide (human connecting peptide) in serum, buffered solution or cell culture medium.
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Mature Connecting Peptide (C-Peptide) is a 31 amino acid peptide that is a product from the proteolytic processing of proinsulin. Through its G-protein coupled membrane receptor, C-Peptide activates Ca2+-dependent intracellular signaling pathways, resulting in subsequent activation of both Na+/K+ ATPase and endothelial nitric oxide synthase. C-Peptide and insulin are co-secreted in equimolar amounts in the bloodstream by pancreatic beta cells of the islets of Langerhans. However, C-Peptide has a longer half-life than insulin in serum, making it a good indicator of endogenous insulin production. A lack of endogenous C-Peptide contributes to development of long-term microvascular complications in the nerves, the kidneys, and the retina in Type 1 diabetes patients. C-Peptide has been considered as a possible therapy to prevent long-term complications of diabetes.
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||Peptide|
|Experimental Type||In vitro|
|Product Brand Name||AlphaLISA|
|Shipping Condition||Blue Ice|
|Unit Size||5,000 assay points|
The aim of this work was to compare the performance of the AlphaLISA® amplified luminescent proximity homogeneous assay technology platform and an alternative time-resolved fluorescenceresonance energy transfer (TR-FRET) technology assay platform for the detection of five biomarkers.
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.