Carbonic anhydrases (CAs), also designated carbonate dehydratases or carbonate hydrolyases, form a large family of genes that encode zinc metalloenzymes of great physiologic importance. As catalysts of the reversible hydration of carbon dioxide, these enzymes participate in a variety of biologic processes, including respiration, acid-base balance, bone resorption and calcification as well as the formation of aqueous humor, cerebrospinal fluid, saliva and gastric acid. Genes in the α-carbonic anhydrase family encode either active carbonic anhydrase isozymes or "acatalytic" (devoid of CO2 hydration activity) carbonic anhydrase-related proteins. Human CA I (CA1) is encoded by the CA1 gene, which maps to a region on chromosome 8 that harbors a cluster of CA genes. CA I localizes to the cytoplasm and research indicates that a severe deficiency of CA I does not result in any obvious hematological or renal consequences.
Background References
1. Briganti F et al. Carbonic anhydrase catalyzes cyanamide hydration to urea: is it mimicking the physiological reaction J Biol Inorg Chem 4:528-536 (1999).
2. Kannan K K et al. Crystal structure of human erythrocyte carbonic anhydrase B. Three-dimensional structure at a nominal 2.2-A resolution. Proc Natl Acad Sci USA 72:51-55 (1975).
Western blot analysis of Carbonic Anhydrase I on different lysates with Rabbit anti-Carbonic Anhydrase I antibody (ET7108-77) at 1/1,000 dilution.
Lane 1: TF-1 cell lysate (20 µg/Lane) Lane 2: K-562 cell lysate (20 µg/Lane) Lane 3: Zebrafish tissue lysate (40 µg/Lane) Lane 4: Human lung tissue lysate (40 µg/Lane)
Predicted band size: 29 kDa Observed band size: 29 kDa
Exposure time: 4 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET7108-77) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Western blot analysis of Carbonic Anhydrase I on different lysates with Rabbit anti-Carbonic Anhydrase I antibody (ET7108-77) at 1/500 dilution.
Lane 1: Mouse colon tissue lysate Lane 2: Rat bone marrow tissue lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 29 kDa Observed band size: 29 kDa
Exposure time: 30 seconds;
12% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET7108-77) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.
Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-Carbonic Anhydrase I antibody (ET7108-77) at 1/800 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7108-77) at 1/800 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded human small intestine tissue using anti-Carbonic Anhydrase I antibody. Counter stained with hematoxylin.
Immunohistochemical analysis of paraffin-embedded mouse spleen tissue using anti-Carbonic Anhydrase I antibody. Counter stained with hematoxylin.
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