Human MAPK/MAK/MRK overlapping kinase (RAGE) ELISA Kit

Tumor antigens recognized by cytolytic T cells (CTLs) can be classified into 4 groups: shared tumor-specific antigens encoded by MAGE-type genes that are silent in most normal tissues except testis and expressed in many tumors; differentiation antigens that are also expressed in normal cells; antigens resulting from tumor-specific point mutations; and antigens overexpressed in tumor cells. By screening cells expressing both HLA-B7 and kidney tumor cell RNA with autologous CTLs, followed by anchored PCR analysis, Gaugler et al. (1996) isolated a cDNA encoding RAGE. RT-PCR analysis detected expression only in retina and in a number of tumor cell types. One putative RAGE open reading frame encodes a 40-amino acid protein and includes the peptide recognized by HLA-B7-restricted CTLs.

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Human MAPK/MAK/MRK overlapping kinase (RAGE) ELISA Kit has high sensitivity and excellent specificity for detection of Human RAGE. No significant cross-reactivity or interference between Human RAGE and analogues was observed.

This Human MAPK/MAK/MRK overlapping kinase (RAGE) ELISA Kit employs a two-site sandwich ELISA to quantitate RAGE in samples. An antibody specific for RAGE has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyRAGE present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for RAGE is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of RAGE bound in the initial step. The color development is stopped and the intensity of the color is measured.

• Do not mix components from different kit lots or use reagents beyond the kit expiration date.• Allow all reagents to warm to room temperature for at least 30 minutes before opening.• Pre-rinse the pipet tip with reagent, use fresh pipet tips for each sample, standard and reagent to avoid contamination.• Unused wells must be kept desiccated at 4 °C in the sealed bag provided.• Mix Thoroughly is very important for the result. It is recommended using low frequency oscillator or slight hand shaking every 10 minutes.• It is recommended that all samples and standards be assayed in duplicate or triplicate.