Human Carbonic Anhydrase VA/CA5A Antibody

Catalog #: AF3049 Datasheet / COA / SDS

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AF3049
AF3049-SP

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Detection of Human Carbonic Anhydrase VA/CA5A by Western Blot.

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human Carbonic Anhydrase VA/CA5A Antibody Summary

Species Reactivity

Human

Specificity

Detects human Carbonic Anhydrase VA/CA5A in direct ELISAs and Western blots. In direct ELISAs, approximately 4% cross-reactivity with recombinant human (rh) CA5B is observed and less than 1% cross-reactivity with rhCA2 and rhCA7.

Source

Polyclonal Sheep IgG

Purification

Antigen Affinity-purified

Immunogen

E. coli-derived recombinant human Carbonic Anhydrase VA/CA5A
Ala40-Ser305
Accession # P35218

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Applications

Recommended Concentration

Sample

Western Blot

1 µg/mL

See below

Immunohistochemistry

5-15 µg/mL

See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot

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Detection of Human Carbonic Anhydrase VA/CA5A by Western Blot. Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line. PVDF Membrane was probed with 1 µg/mL of Goat Anti-Human Carbonic Anhydrase VA/CA5A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3049) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for Carbonic Anhydrase VA/CA5A at approximately 33 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.

Immunohistochemistry

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Carbonic Anhydrase VA/CA5A in Human Liver. Carbonic Anhydrase VA/CA5A was detected in immersion fixed paraffin-embedded sections of human liver using Sheep Anti-Human Carbonic Anhydrase VA/CA5A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3049) at 10 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to hepatocyte cytoplasm. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Sterile PBS to a final concentration of 0.2 mg/mL.

Reconstitution Buffer Available

Reconstitution Buffer 1 (PBS)

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: Carbonic Anhydrase VA/CA5A

Carbonic Anhydrase catalyzes the reversible reaction of CO₂ + H₂O = HCO₃^- + H⁺, which is fundamental to many processes such as respiration, renal tubular acidification and bone resorption (1). Topics in a CA meeting (6^th International Conference on the CAs, June 20 - 25, 2003, Slovakia) ranged from the use of CAs as markers for tumor and hypoxia in the clinic, as a nutritional supplement in milk, and as a tool for CO₂ removal and mosquito control in industry. Carbonic anhydrase VA encoded by the CA5A gene is a mitochondrial protein (2, 3). In comparison with another mitochondrial CA (CA5B), CA5A has different tissue distribution and chromosomal location (4, 5). Expression and inhibitor studies of different CAs in the rat pancreatic beta cells indicate that CA5A may be involved in the regulation of insulin secretion (6). CA5A may also participate in the detoxification of ammonia produced in the gastrointestinal tract by providing bicarbonate to carbamyl phosphate synthetase I (7). The amino acid sequence of recombinant human CA5A (residues 40 to 305) is 79%, 77%, and 76% identical to that of canine, bovine, and rat/mouse.

References

Hewett-Emmett, D. and R.E. Tashian (1996) Mol. Phylogenet. Evol. 5:50.
Nagao, Y. et al. (1993) Proc. Natl. Acad. Sci. USA 90:7623.
Nagao, Y. et al. (1995) Genomics 28:477.
Shah, G.N. et al. (2000) Proc. Natl. Acad. Sci. USA 97:1677.
Fujikawa-Adachi, K. et al. (1999) J. Biol. Chem. 274:21228.
Parkkila, A.K. et al. (1998) J. Biol. Chem. 273:24620.
Saarnio, J. et al. (1999) J. Histochem. Cytochem. 47:517.

Entrez Gene IDs

763 (Human); 54233 (Rat)

Alternate Names

CA5A; CA5carbonic anhydrase 5A, mitochondrial; Carbonate dehydratase VA; carbonic anhydrase V, mitochondrial; Carbonic Anhydrase VA; carbonic anhydrase VA, mitochondrial; carbonic dehydratase; CAV; CAVA; CA-VA; EC 4.2.1.1