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EK000123-EK0757: Human Fetuin A ELISA Kit
Range 31.2pg/ml-2000pg/ml
Sensitivity < 2pg/ml
Specificity: No detectable cross-reactivity with any other protein.
Application: For quantitative detection of Human Fetuin A in tissue lysates or cell culture supernates. The significance of detection in sera, plasma, body fluids is undetermined.
Expiration: Four months at 4°C and eight months at -20°C.
Background
Alpha2-HS glycoprotein (AHSG), also known as fetuin-A, is a plasma protein displaying high-affinity interaction with calcium phosphate, by which ectopic vascular calcification is prevented. The AHSG polymorphism is attributable to the hereditary variation of AHSG and phosphate serum levels, which may affect skeletal development and chronic disorders such as vascular calcification.1Human plasma protein alpha 2-HS-glycoprotein (AHSG) is composed of two polypeptide chains, A and B, encoded by a single mRNA. Southern blot analysis of mouse x human somatic cell hybrids has mapped the AHSG gene to human chromosome 3 in the region 3q21----qter (Lee et al., 1987). Using a recombinant plasmid containing a 1,538 bp insert spanning the entire AHSG coding region, AHSG was localized to chromosomal bands 3q27----q29 by in situ hybridization.2
Principle
Fetuin A ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. Human Fetuin A specific-specific polyclonal antibodies were precoated onto 96-well plates. The human specific detection polyclonal antibodies were biotinylated. The test samples and biotinylated detection antibodies were added to the wells subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the Human Fetuin A amount of sample captured in plate.
Reference
1. Osawa, M., Tian, W., Horiuchi, H., Kaneko, M., Umetsu, K. Association of alpha-2-HS glycoprotein (AHSG, fetuin-A) polymorphism with AHSG and phosphate serum levels. Hum. Genet. 116: 146-151, 2005.
2. Magnuson, V. L., McCombs, J. L., Lee, C.-C., Yang, F., Bowman, B. H., McGill, J. R. Human alpha-2-HS-glycoprotein localized to 3q27-q29 by in situ hybridization. Cytogenet. Cell Genet. 47: 72-74, 1988.
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