The AlphaLISA® immunoassay kit for detection and quantitation of human VEGF (VEGF-A) in serum, cell culture supernatants, and other sample types allows for fast, reproducible, and sensitive detection without the need for time-consuming wash steps or complicated assay development.
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The antibodies in this kit recognize VEGF-A (VEGF165 or isoform d), VEGF162, VEGF145 (isoform e) and VEGF121 (isoform f). One of the sandwiching antibodies is a mouse monoclonal, clone 26503. The other sandwiching antibody is a goat polyclonal raised against human recombinant VEGF165.
Vascular endothelial growth factor (VEGF-A or VEGF) is a homodimeric 34-42 kDa heparin-binding glycoprotein specific for endothelial cells. Currently at least eight variants of VEGF generated by alternative splicing of the same gene have been identified. The various isoforms have different heparin- and neuropilin(-1 or -2)-binding properties as well as solubility characteristics reflecting the different functional properties of the VEGF forms. VEGF is believed to play important roles in inflammation and during normal and pathological angiogenesis, a process that is associated with wound healing, embryonic development, and growth and metastasis of solid tumors. Elevated levels of VEGF have been observed in synovial fluids of rheumatoid arthritis patients and sera of cancer patients.
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||500 assay points|
For the detection of three biomarkers in complex sample matrices, the AlphaLISA and Electrochemiluminescent (ECL) assay technologies were shown to have similar: Assay windows (linear dynamic range), Lower and upper detection limit, Intra-and inter-assay precision (lower % CV) The advantages of using AlphaLISA over ECL are: Shorter total assay duration No wash steps No shaking Lower sample volume requirement for equivalent performance Less expensive instrument and plates required
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.