Carbonic Anhydrase XII/CA12 Antibody [Unconjugated]

Western blot shows lysates of human kidney tissue and A549 human lung carcinoma cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human Carbonic Anhydrase XII/CA12 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2190) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Carbonic Anhydrase XII/CA12 at approximately 45-50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Simple Western lane view shows lysates of A549 human lung carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for Carbonic Anhydrase XII/CA12 at approximately 60 kDa (as indicated) using 10 µg/mL of Goat Anti-Human Carbonic Anhydrase XII/CA12 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2190) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Differential expression of CAIX and CAXII in patient-derived tumor grafts. Panel A. Frozen tissue samples from PDX tumors were homogenized in RIPA buffer containing protease inhibitor. Western blots were probed with antibodies for CAIX, CAXII, CAII, ER, E-cadherin, actin, and GAPDH. Panel B. Tumor growth rates of orthotopically implanted, cryo-preserved tumor tissue was evaluated in NOD/SCID mice. Panel C. Immunohistochemistry was utilized to evaluate expression of CAIX, CAXII, ER, and Ki67 in TNBC (HCl-001) and ER/PR-positive (HCl-011) tumors from Panel B. Magnification: primary objective magnification, 10x. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/29965974), licensed under a CC-BY license. Not internally tested by R&D Systems.

Differential expression of CAs in TNBC and luminal breast cancer cells. Cell were grown to 70% confluence and then exposed to normoxic (N) or 1% oxygen (H), hypoxic conditions. After 16 h, cells were washed with PBS and extracted in RIPA buffer containing protease inhibitors. Equal protein was loaded onto SDS PAGE gels, and then transferred to nitrocellulose for western blot analysis. Panel A. CA expression in TNBC cells: UFH = UFH-001, MDA = MDA-MB-231-LM2, HBL = HBL-100, S159 = Sum 159, BT = BT-549 cells. Panel B. CA expression in luminal breast cancer cells: T47D, MCF7 = MCF-7, SKBR = SKBR-3, S52 = SUM-52 cells. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/29965974), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: WB, Simple Western
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

Carbonic Anhydrase XII/CA12 Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human Carbonic Anhydrase XII/CA12
  Ala25-Gln291
  Accession # O43570
• Specificity: Detects human Carbonic Anhydrase XII/CA12 in direct ELISAs and Western blots. In Western blots, approximately 40% cross-reactivity with recombinant mouse Carbonic Anhydrase XII/CA12 is observed and less than 5% cross-reactivity with recombinant human CA1, 3, 5A, 5B, 6, 7, 8, and 10 is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: CA12
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Simple Western 10 ug/mL
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  12 months from date of receipt, -20 to -70 degreesC as supplied. 1 month, 2 to 8 degreesC under sterile conditions after reconstitution. 6 months, -20 to -70 degreesC under sterile conditions after reconstitution.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Carbonic Anhydrase XII/CA12 Antibody [Unconjugated]

• CA12
• Carbonate dehydratase XII
• carbonic anhydrase 12
• Carbonic Anhydrase XII
• carbonic anhydrase XIICAXII
• carbonic dehydratase
• CA-XII
• EC 4.2.1.1
• FLJ20151
• HsT18816
• Tumor antigen HOM-RCC-3.1.3

Background

Carbonic Anhydrase (CA) catalyzes the reversible reaction of CO 2 + H 2 O = HCO 3 - + 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 CO 2 removal and mosquito control in industry. CA12 is a type I membrane enzyme highly expressed in colon, kidney, prostate, intestine and activated lymphocytes and moderately expressed in pancreas, ovary, and testis (2, 3). It is over-expressed in some renal cell cancers and in glaucoma (2, 4). Two alternatively spliced forms exist, which either contains or lacks a 11 amino acid segment just before the transmembrane domain (5). The secreted, purified recombinant human CA12 contains the residues that are common for both forms.

1. Hewett-Emmett, D. and R.E. Tashian (1996) Mol. Phylogenet. Evol. 5:50.
2. Tuereci, O. et al. (1998) Proc. Natl. Acad. Sci. USA 95:7608.
3. Ivanov, S.V. et al. (1998) Proc. Natl. Acad. Sci. USA 95:12596.
4. Liao, S.-Y. et al. (2003) J. Med. Genet. 40:257.
5. Strausberg, R.L. et al. (2002) Proc. Natl. Acad. Sci. USA 99:16899.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Carbonic Anhydrase XII/CA12 Antibody (AF2190)(2)

Publications using AF2190

• Chen Z, Ai L, Mboge MY et al. Differential expression and function of CAIX and CAXII in breast cancer: A comparison between tumorgraft models and cells. PLoS ONE. 7/2/2018 [PMID: 29965974]
• McIntyre A, Patiar S, Wigfield S Carbonic anhydrase IX promotes tumor growth and necrosis in vivo and inhibition enhances anti-VEGF therapy. Clin. Cancer Res., 2012-04-12;18(11):3100-11. 2012-04-12 [PMID: 22498007] (Western Blot, Human)

Bioinformatics

• Gene Symbol: CA12
• Uniprot:
  • O43570()