Human Dolichyl-diphosphooligosaccharide--protein glycosyltransferase subunit DAD1 (DAD1) ELISA Kit

DAD1, the defender against apoptotic cell death, was initially identified as a negative regulator of programmed cell death in the temperature sensitive tsBN7 cell line. The DAD1 protein disappeared in temperature-sensitive cells following a shift to the nonpermissive temperature, suggesting that loss of the DAD1 protein triggered apoptosis. DAD1 is believed to be a tightly associated subunit of oligosaccharyltransferase both in the intact membrane and in the purified enzyme, thus reflecting the essential nature of N-linked glycosylation in eukaryotes.

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Human Dolichyl-diphosphooligosaccharide--protein glycosyltransferase subunit DAD1 (DAD1) ELISA Kit has high sensitivity and excellent specificity for detection of Human DAD1. No significant cross-reactivity or interference between Human DAD1 and analogues was observed.

This Human Dolichyl-diphosphooligosaccharide--protein glycosyltransferase subunit DAD1 (DAD1) ELISA Kit employs a two-site sandwich ELISA to quantitate DAD1 in samples. An antibody specific for DAD1 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyDAD1 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for DAD1 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 DAD1 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.