Human Rac GTPase-activating protein 1 (RACGAP1) ELISA Kit

The predicted 527-amino acid RACGAP1 protein has a large N-terminal region containing a protein kinase C -like cysteine-rich motif. The authors determined that RACGAP1 shares highest homology with the Drosophila RnRacGAP and the chimerins of rat and human. Functional analysis showed that the GAP domain of RACGAP1 exhibits strong GAP activity towards CDC42, RAC1, and RAC2. Northern blot analysis detected an approximately 3.2-kb RACGAP1 transcript that was most abundantly expressed in testis, with low expression in most other tissues. Western blot analysis detected a RACGAP1 protein of 58 kD in testis extracts. In situ hybridization showed that RACGAP1 expression is restricted to germ cells in mature testis.

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Human Rac GTPase-activating protein 1 (RACGAP1) ELISA Kit has high sensitivity and excellent specificity for detection of Human RACGAP1. No significant cross-reactivity or interference between Human RACGAP1 and analogues was observed.

This Human Rac GTPase-activating protein 1 (RACGAP1) ELISA Kit employs a two-site sandwich ELISA to quantitate RACGAP1 in samples. An antibody specific for RACGAP1 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyRACGAP1 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for RACGAP1 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 RACGAP1 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.