||Recombinant human CA2 was expressed in E.coli and purified by using conventional chromatography, 29.2 kDa (260aa)
||Carbonic anhydrase 2 (CA2) is one of fourteen forms of humanα carbonic anhydrases. Carbonic anhydrase catalyzes reversible hydration of carbon dioxide. CA2 is a cytosolic enzyme with the highest activity among all known CAs. Mutations in the CA2 gene result in the CA2 deficiency syndrome, an autosomal recessive disorder that produces osteopetrosis, renal tubular acidosis and cerebral calcification.
||> 95% by SDS - PAGE
|Sequences of amino acids:
||MSHHWGYGKH NGPEHWHKDF PIAKGERQSP VDIDTHTAKY DPSLKPLSVS YDQATSLRIL NNGHAFNVEF DDSQDKAVLK GGPLDGTYRL IQFHFHWGSL DGQGSEHTVD KKKYAAELHL VHWNTKYGDF GKAVQQPDGL AVLGIFLKVG SAKPGLQKVV DVLDSIKTKG KSADFTNFDP RGLLPESLDY WTYPGSLTTP PLLECVTWIV LKEPISVSSE QVLKFRKLNF NGEGEPEELM VDNWRPAQPL KNRQIKASFK
||1 mg/ml (determined by Bradford assay)
||Liquid. In 20 mM Tris-HCl buffer (pH 8.0) containing 1mM DTT, 50mM NaCl, 10% glycerol
||Specific activity is 50-70 nmoles/min/ug and was obtained by measuring the increase in the amount of p-nitrophenol by its esterase activity. Specific activity is defined as the amount of p-nitrophenol that 1ug of enzyme can reduce at 25℃ for 1 minute.
||1. Prepare assay buffer into a suitable container: The final concentrations are 12.5 mM Tris, 75 mM NaCl, pH 7.5. 2. Dilute 4-Nitrophenyl-Acetate(4-NPA) to 2 mM in Assay Buffer. 3. Dilute recombinant CA2 protein with various concentrations (1ug, 2ug, 5ug) in 50 ul assay buffer. 4. Load 50 μL of 2mM 4-NPA into diluted recombinant CA2 solutions, and start the reaction by adding 50 μL of 2 mM Substrate. 5. Read at a wavelength of 405 nm (bottom read) in kinetic mode for 5 minutes.
||Can be stored at +4°C short term (1-2 weeks). For long term storage, aliquot and store at -20°C or -70°C. Avoid repeated freezing and thawing cycles.