Human Ubiquitin D (UBD) ELISA Kit

Ubiquitin D is a protein encoded by the UBD gene.Using selective cDNA hybridization of a YAC containing the HLA-F locus region on chromosome 6, followed by Southern and Northern blot analyses, Fan et al. (1996) isolated a cDNA, which they called 1F12, encoding a protein similar to diubiquitin. The 1.1-kb transcript was detected only in B-cell lines transformed by Epstein-Barr virus. By subtractive hybridization of dendritic cell (DC) libraries and EST database searching, Bates et al. (1997) obtained a full-length cDNA encoding diubiquitin. The predicted 165-amino acid protein has 2 ubiquitin-like domains, each of which has 2 cysteines. Southern blot analysis indicated that the cDNA represents a single-copy gene.

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

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