Detects mouse Carbonic Anhydrase IV/CA4 in direct ELISAs and Western blots. In direct ELISAs and Western blots, approximately 15% cross‑reactivity with recombinant human (rh) CA4 is observed and less than 2% cross-reactivity with rhCA3, rhCA10, rhCA13, recombinant mouse (rm) CA9, and rmCA12 is observed.
Detection of Mouse Carbonic Anhydrase IV/CA4 by Western Blot.
Western blot shows lysates of mouse kidney tissue. PVDF membrane was probed with 0.25 µg/mL of Goat Anti-Mouse Carbonic Anhydrase IV/CA4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2414) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for Carbonic Anhydrase IV/CA4 at approximately 35 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Carbonic Anhydrase IV/CA4 in Mouse Kidney.
Carbonic Anhydrase IV/CA4 was detected in perfusion fixed frozen sections of mouse kidney using Goat Anti-Mouse Carbonic Anhydrase IV/CA4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2414) at 3 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in epithelial cells. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.
Detection of Mouse Carbonic Anhydrase IV/CA4 by Simple WesternTM.
Simple Western lane view shows lysates of mouse lung tissue and mouse kidney tissue, loaded at 0.2 mg/mL. A specific band was detected for Carbonic Anhydrase IV/CA4 at approximately 51-52 kDa (as indicated) using 12.5 µg/mL of Goat Anti-Mouse Carbonic Anhydrase IV/CA4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2414) 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.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
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 IV/CA4
Carbonic anhydrase (CA) 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 (6th International Conference on the CAs, June 20‑25, 2003, Slovakia) ranged from use of CAs as markers for tumor and hypoxia in clinic, as nutritional supplement in milk, and as a tool for CO2 removal and mosquito control in industry. The deduced amino acid sequence of mouse CA4 consists of a signal peptide (residues 1‑17), an ectodomain (residues 18‑277) and a pro peptide (residues 278‑305), which is removed in the mature form (2). The native enzyme is attached to the membrane by a GPI‑anchor. Recombinant mouse CA4 corresponds to the ectodomain and exhibits the activity as described in Activity Assay Protocol.
Hewett-Emmett, D. and R.E. Tashian (1996) Mol. Phylogenet. Evol. 5:50.
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