Human Ras-related protein Rap-1b (RAP1B) ELISA Kit

The deduced 184-amino acid protein is 95% identical to RAP1A. RAP1B shares several properties with the RAS protein, suggesting that it may bind GTP/GDP and have a membrane location.RAP1B is mainly located at the membrane, but translocates to the cytosol upon activation. In activated platelets, RAP1B interacts with the reorganized actin-based cytoskeleton. RAP1B is activated by phosphorylation, increased intracellular Ca (2+), and by agonist-induced stimulation of Gi, which results in the rapid binding of GTP to RAP1B. RAP1B is posttranslationally modified by the geranylgeranylation of cys181. Further modifications cause proteolytic removal of the last 3 C-terminal amino acids, followed by partial methylation of the remaining terminal cysteinyl carboxyl group.

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Human Ras-related protein Rap-1b (RAP1B) ELISA Kit has high sensitivity and excellent specificity for detection of Human RAP1B. No significant cross-reactivity or interference between Human RAP1B and analogues was observed.

This Human Ras-related protein Rap-1b (RAP1B) ELISA Kit employs a two-site sandwich ELISA to quantitate RAP1B in samples. An antibody specific for RAP1B has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyRAP1B present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for RAP1B 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 RAP1B 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.