Rat Ubiquitin (Ub) ELISA Kit

Posttranslational modification of proteins by the addition of the small protein SUMO, or sumoylation, regulates protein structure and intracellular localization. SAE1 and UBA2 form a heterodimer that functions as a SUMO-activating enzyme for the sumoylation of proteins. The deduced SAE1 and SAE2 proteins contain 347 and 640 amino acids, respectively. SAE1 shares sequence similarity with the N terminus of ubiquitin-activating E1 enzymes, and SAE2 share sequence similarity with the C terminus of E1 enzymes. Both SAE subunits contain a conserved nucleotide-binding motif, and SAE2 contains an E1-like active-site cysteine. SAE2 has a calculated molecular mass of 72 kD. It had an apparent molecular mass of 90 kD by SDS-PAGE.

The product listed herein is for research use only and is not intended for use in human or clinical diagnosis. Suggested applications of our products are not recommendations to use our products in violation of any patent or as a license. We cannot be responsible for patent infringements or other violations that may occur with the use of this product.

Rat Ubiquitin (Ub) ELISA Kit has high sensitivity and excellent specificity for detection of Rat Ub. No significant cross-reactivity or interference between Rat Ub and analogues was observed.

This Rat Ubiquitin (Ub) ELISA Kit employs a two-site sandwich ELISA to quantitate Ub in samples. An antibody specific for Ub has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyUb present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for Ub 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 Ub 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.