Fetuin A ELISA Kit (Human) (OKBB00323)

Catalog No: OKBB00323 (Formerly GWB-ZZD177)

Size:96 Tests

Human AHSG ELISA Kit standard curve

Product Info

• Predicted Species Reactivity: Human
• Application: ELISA
• ELISA Kit Detection Method: Colorimetric, OD450 nm
• ELISA Kit Duration: ~ 3 Hours
• ELISA Kit Principle: Aviva's human Fetuin A ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for Fetuin A has been precoated onto 96-well plates. Standards(NS0, A19-V367) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for Fetuin A is added 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 absorbance of yellow is proportional to the human Fetuin A amount of sample captured in plate.
• ELISA Kit Range: 0.78 ng/ml - 50 ng/ml
• ELISA Kit Reproducibility:

  	Intra-Assay Precision			Inter-Assay Precision
  Sample	1	2	3	1	2	3
  Mean (ng/mL)	2.17	8.32	12.55	2.39	8.75	13.64
  St. Dev.	0.089	0.441	0.602	0.129	0.543	0.777
  %CV	4.10	5.30	4.79	5.39	6.20	5.69

• ELISA Kit Component:
  • Lyophilized recombinant mouse C-MET standard: 10 ng/tube x 2
  • Anti-mouse C-MET Antibody Well Plate: 96 Wells
  • Sample diluent buffer: 30 mL
  • Biotinylated anti-mouse C-MET antibody: 130 uL, dilution 1:100
  • Antibody diluent buffer: 12 mL
  • Avidin-Biotin-Peroxidase Complex (ABC): 130 uL, dilution 1:100
  • ABC diluent buffer: 12 mL
  • TMB color developing agent: 10 mL
  • TMB stop solution: 10 mL
• Additional Information: Range: 0.312ng/ml => 20ng/ml
  Principle: Aviva’s Human 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.
  Notes: 1. To inspect the validity of experiment operation and the appropriateness of sample dilution proportion, pilot experiment using standards and a small number of samples is recommended.
  2. The TMB Color Developing agent is colorless and transparent before using, contact us freely if it is not the case.
  3. Before using the Kit, spin tubes and bring down all components to the bottom of tubes.
  4. Duplicate well assay is recommended for both standard and sample testing.
  5. Don’t let 96-well plate dry, for dry plate will inactivate active components on plate.
  6. Don’t reuse tips and tubes to avoid cross contamination.
  7. To avoid to use the reagents from different batches together.
  8. In order to avoid marginal effect of plate incubation due to temperature difference ( reaction may be stronger in the marginal wells), it is suggested that the diluted ABC and TMB solution will be pre-warmed in 37C for 30 min before using.
  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
• Reconstitution and Storage: Store at 4C for 6 months, at -20C for 12 months. Avoid multiple freeze-thaw cycles (Shipped with wet ice.)
• Sample Type: cell culture supernates, serum or plasma (heparin, EDTA)
• Sensitivity: <10 pg/ml
• Predicted Homology Based on Immunogen Sequence: No detectable cross-reactivity with any other cytokine.
• Reference: 11. 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.
• Specificity: Natural and recombinant human Fetuin A
• Immunogen: Expression system for standard: NS0; Immunogen sequence: A19-V367
• Description: For quantitative detection of human Fetuin A in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).

Target Info

• Gene Symbol: AHSG
• Gene Full Name: alpha-2-HS-glycoprotein
• Alias Symbols: A2HS, AHS, Ahsg, Alpha 2 HS glycoprotein, Alpha-2-HS-glycoprotein chain B, Alpha-2-Z-globulin, Ba-alpha-2-glycoprotein, FETUA, FETUA_HUMAN, Fetuin-A, HSGA
• NCBI Gene Id: 197
• Protein Name: Alpha-2-HS-glycoprotein
• Description of Target: 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. 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
• Uniprot ID: P02765
• Protein Accession #: NP_001613.2
• Nucleotide Accession #: NM_001622.2