Zinc-Alpha-2-Glycoprotein Human ELISA

Regulatery Status: RUO
Type: Sandwich ELISA, HRP-labelled antibody
Other Names Status: ZA2G, ZAG, AZGP1, Zn-alpha-2-glycoprotein, Zn-alpha-2-GP, ZNGP1
Species: Human
Catalog No Size
Product Catalog No: RD191093100R Pack Size: 96 wells (1 kit)

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Product Features

Zinc-alpha-2-glycoprotein (ZAG, ZA2G, Azgp1, ZNGP1, Lipid-Mobilizing Factor, LMF) is a soluble 41 kDa glycoprotein belonging to the immunoglobuline protein family and consisting of a single polypeptide chain. Human ZAG shares 59% sequence identity with the murine homolog.

ZAG is closely related to antigens of the class1 major histocompatibility complex (MHC I) and shares 30-40 % sequence identity with the heavy chain of MHC I. Most MHC-I members heterodimerize with beta-2-microglobuline (b2m) and bind peptides derived from intracellular proteins to present them to cytotoxic T cells. In contrast, ZAG is a soluble protein rather than being anchored to plasma membranes that acts independently on b2m and binds the hydrophobic ligand which may relate to its function in lipid metabolism.

ZAG is widespread in body fluids and is also found in various human tissues such as adipose tissue, prostate, breast, skin, salivary gland, trachea, broncheus, lung, gastrointestinal tract, pancreas, liver and kidney. ZAG acts as a lipid mobilizing factor to induce lipolysis in adipocytes and plays an important role in lipid utilization and loss of adipose tissue, especially during cachexia, which occurs in patient suffering from cancer, AIDS and other chronic illnesses. The role of ZAG in cancer cachexia is also connected with its ability to directly influence expression of uncoupling proteins (UCPs) which are implicated in the regulation of energy balance. In human adipocytes, ZAG expression is regulated particularly through TNF-alpha and the PPAR gamma nuclear receptor. ZAG expression is also upregulated by glucocorticoides and attenuated by eicosapentaenoic acid (EPA) and beta-3-adrenoreceptor antagonists.

ZAG is overexpressed in certain human malignant tumors such as prostate, breast, lung or bladder cancer and can relate to tumor differentiation. Additionally, ZAG plays a role in obesity, diabetic kidney disorders, frontotemporal dementia and regulation of melanin production by melanocytes.

ZAG is proposed to have a therapeutic use in obesity and cachexia. It can be used as a marker for clinical analysis of diabetic nephropathy and as a marker for certain tumors..

Technical Sheet / Info

Intended use

The RD191093100R Human Zinc-Alpha-2-Glycoprotein ELISA is a sandwich enzyme immunoassay for the quantitative measurement of human zinc-alpha-2-glycoprotein in serum and plasma.

  • The kit measures zinc-alpha-2-glycoprotein in serum and plasma (EDTA, citrate, heparin)
  • Assay format is 96 wells
  • Quality Controls are human serum based. No animal sera are used
  • Standard is recombinant protein based
  • Components of the kit are provided ready to use, concentrated or lyophilized

Clinical Application

  • Energy metabolism and body weight regulation
  • Oncology

Test principle

In the BioVendor Human Zinc-Alpha-2-Glycoprotein (ZA2G) ELISA, standards, quality controls and samples are incubated in microplate wells pre-coated with polyclonal anti-human ZA2G antibody. After 60 minutes incubation and washing, polyclonal anti-human ZA2G antibody, conjugated with horseradish peroxidase (HRP) is added to the wells and incubated for 60 minutes with captured ZA2G. Following another washing step, the remaining HRP conjugate is allowed to react with the substrate solution (TMB). The reaction is stopped by addition of acidic solution and absorbance of the resulting yellow product is measured. The absorbance is proportional to the concentration of ZA2G. A standard curve is constructed by plotting absorbance values against concentrations of standards, and concentrations of unknown samples are determined using this standard curve.

Summary of protocol

  • Prepare Dilution Buffer
  • Reconstitute QCs and Master Standard and prepare set of standards
  • Dilute samples 5 000×
  • Add 100 μl Standards, QCs and samples
  • Incubate at RT for 1 hour/300 rpm
  • Wash plate 3 times
  • Add 100 μl Conjugate Solution
  • Incubate at RT for 1 hour/300 rpm
  • Wash plate 3 times
  • Add 100 μl Substrate Solution
  • Incubate at RT for 10 min
  • Add 100 μl Stop Solution
  • Read absorbance and calculate results

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References

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