Human N-acetylglucosaminyl-phosphatidylinositol de-N-acetylase (PIGL) ELISA Kit

Glycosylphosphatidylinositol (GPI) is used as a membrane anchor by many eukaryotic cell surface proteins. The first step in GPI biosynthesis involves the transfer of N-acetylglucosamine (GlcNAc) from UDP-GlcNAc to phosphatidylinositol (PI) . The second step is de-N-acetylation of GlcNAc-PI. PIGL encodes a deduced 252-amino acid protein that shares 77% identity with the rat protein. Using transfection into mammalian PIGL-deficient cells, Watanabe et al. (1999) demonstrated that S. cerevisiae Gpi12 is the ortholog of human and rat PIGL. Disruption of Gpi12 in yeast resulted in a lethal phenotype. Using purified, recombinant rat PIGL, Watanabe et al. (1999) demonstrated that PIGL has GlcNAc-PI de-N-acetylase activity in vitro, which is enhanced by metal ions.

PIGL; N-acetylglucosaminyl-phosphatidylinositol de-N-acetylase; N-acetylglucosaminylphosphatidylinositol deacetylase; phosphatidylinositol glycan; class L

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.

Human N-acetylglucosaminyl-phosphatidylinositol de-N-acetylase (PIGL) ELISA Kit has high sensitivity and excellent specificity for detection of Human PIGL. No significant cross-reactivity or interference between Human PIGL and analogues was observed.

This Human N-acetylglucosaminyl-phosphatidylinositol de-N-acetylase (PIGL) ELISA Kit employs a two-site sandwich ELISA to quantitate PIGL in samples. An antibody specific for PIGL has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyPIGL present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for PIGL 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 PIGL bound in the initial step. The color development is stopped and the intensity of the color is measured.

PIGL; N-acetylglucosaminyl-phosphatidylinositol de-N-acetylase; N-acetylglucosaminylphosphatidylinositol deacetylase; phosphatidylinositol glycan; class L

• 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.