Mouse Trehalase (TREH) ELISA Kit

Trehalose (1-alpha-D-glucopyranosyl alpha-D-glucopyranoside) is widely distributed in nature, being the storage disaccharide of fungi and the blood sugar of insects. Trehalase (EC 3.2.1.28) is an intrinsic glycoprotein of the small intestine and renal brush border membranes that hydrolyzes trehalose to 2 glucose molecules. The deduced 583-amino acid protein has a calculated molecular mass of 66.6 kD. It has an N-terminal signal peptide, 5 potential N-glycosylation sites, and a C-terminal hydrophobic region for glycosylphosphatidylinositol (GPI) attachment. Northern blot analysis detected a 2.0-kb transcript mainly in kidney, liver, and small intestine.The trehalase signature sequence from amino acids 162 to 175 of mouse Treh shares 100% identity with the corresponding region in human TREH.

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

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