More info
Assay Range | 15.6-1,000 pg/mL |
Sensitivity | 10.0 pg/mL |
Size | 96T |
Storage | Store at 2 - 8ºC. Keep reconstituted standard and detection Ab at -20 ºC |
Assay Principle | Sandwich ELISA |
Sample volume | 100 µL final volume, dilution factor varies on samples |
Detection Method | Chromogenic |
Kit Components
1. Recombinant Human CA-9 standard: 2 vials
2. One 96-well plate precoated with anti-Human CA-9 Ab
3. Sample diluent buffer: 12 mL - 1
4. Detection antibody: 130 µL, dilution 1:100
5. Streptavidin-HRP: 130 µL, dilution 1:100
6. Antibody diluent buffer: 12 mL x1
7. Streptavidin-HRP diluent buffer: 12 mL x1
8. TMB developing agent: 10 mL x1
9. Stop solution: 10 mL x1.
10. Washing solution (20x): 25 mL x1.
Background
Carbonic Anhydrase IX (CA-9 or CA IX), also known as membrane antigen MN, Renal cell carcinoma-associated antigen G250, or Carbonate dehydratase IX, is a member of the carbonic anhydrase protein family. CA-9 is composed of an N-terminal signal peptide, an extracellular proteoglycan-related domain with a catalytic fragment, a transmembrane segment, and a C-terminal intracellular tail. The ectodomain of CA-9 can be released into the extracellular milieu, which appears to be regulated by the metalloprotease and TNFα-converting enzyme (TACE/ADAM17). This soluble form of CA-9 has been detected in cell culture supernatants, serum and urine. CA-9 plays a major role in regulating pH by catalyzing the reversible hydration of carbon dioxide to carbonic acid, which subsequently decomposes to HCO3- and H3O+. Primarily expressed in the gut, CA-9 is ectopically up-regulated in many types of tumors, such as renal cell carcinoma, non-small cell lung cancer, breast cancer, and cervical cancer. Under hypoxic conditions, the CA-9 gene is induced by its upstream regulator, the hypoxia-inducible factor (HIF)-1 transcription factor. CA-9 facilitates the tumor cells to create an acidified local environment to promote growth and metastasis. Loss of the tumor suppressor gene, von Hippel-Lindau (VHL) also contributes to the overexpression of CA-9 in tumors. It is reported that mutations in the VHL gene can lead to overexpression of CA-9 in cancer cell lines. In addition, CA-9 may participate in regulation of some signaling pathway. The tyrosine moiety of the intracellular tail of CA-9 can be phosphorylated in an epidermal growth factor-dependent manner and the phosphorylated CA-9 can interact with PI-3-kinase.