Carbonic anhydrase 9 (CA9/CAIX) is an enzyme that in humans is encoded by the CA9 gene. Carbonic anhydrases (CAs) are a large family of zinc metalloenzymes that catalyze the reversible hydration of carbon dioxide. They participate in a variety of biological processes, including respiration, calcification, acid-base balance, bone resorption, and the formation of aqueous humor, cerebrospinal fluid, saliva, and gastric acid. This gene is mapped to 9p13.3. CA IX is a transmembrane protein and the only tumor-associated carbonic anhydrase isoenzyme known. It is expressed in all clear-cell renal cell carcinoma, but is not detected in normal kidney or most other normal tissues. It may be involved in cell proliferation and transformation. Sample Type: Cell lysates, cell culture supernates, serum, plasma (Heparin, EDTA or Citrate). Intended Use: Sandwich High Sensitivity BioAssay(TM) ELISA kit for Quantitative Detection of Mouse CA9/Ca Ix. Sensitivity: <10pg/ml Range: 15.6pg/ml-1000pg/ml Specificity: Recognizes natural and recombinant Mouse CA9/Ca Ix. There is no detectable cross-reactivity with other relevant proteins. Test Principle: This Mouse CA9/Ca Ix BioAssay(TM) ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from rat specific for CA9/Ca Ix has been precoated onto 96-well plates. Standards(Expression system for standard: NSO, Immunogen sequence: Q32-D390) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for CA9/Ca Ix 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 density of yellow is proportional to the Mouse CA9/Ca Ix amount of sample captured in plate. Kit Components: 96-well plate precoated with anti-Mouse Ca9 antibody, 1x plate Lyophilized recombinant Mouse Ca9 standard, 2x10ng/tube Biotinylated anti-Mouse Ca9 antibody, 1x130ul (dilution 1:100) Avidin-Biotin-Peroxidase Complex (ABC), 1x130ul (dilution 1:100) Sample diluent buffer, 1x30ml Antibody diluent buffer, 1x12ml ABC diluent buffer, 1x12ml TMB color developing agent, 1x10ml TMB stop solution, 1x10ml PBS washing buffer Powder for 1000ml, 1x buffer Storage and Stability: Store powder at 4C liquid at -20C. Store other components at 4C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
