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 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. CA4 is a GPI‑anchored membrane enzyme expressed on the luminal surfaces of pulmonary (and certain other) capillaries and of proximal renal tubules. It functions as the principal CO2 taste sensor (2). In addition, a genetic mutation (Arg 14 to Trp in the signal peptide) of CA4 was found to cosegregate with the RP17 form of retinitis pigmentosa in two large families and was not found in 36 unaffected family members or 100 controls (3).
Human Carbonic Anhydrase IV/CA4 Antibody
R&D Systems | Catalog # AF2186
Key Product Details
Species Reactivity
Validated:
Human
Cited:
Human
Applications
Validated:
Immunohistochemistry, Western Blot, Simple Western, Immunoprecipitation
Cited:
Western Blot, Immunocytochemistry
Label
Unconjugated
Antibody Source
Polyclonal Goat IgG
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Product Specifications
Immunogen
Mouse myeloma cell line NS0-derived recombinant human Carbonic Anhydrase IV/CA4
Ala19-Lys283
Accession # P22748
Ala19-Lys283
Accession # P22748
Specificity
Detects human Carbonic Anhydrase IV/CA4 in direct ELISAs and Western blots. In direct ELISAs and Western blots, approximately 20% cross‑reactivity with recombinant mouse CA4 is observed and less than 5% cross-reactivity with recombinant human CA1, 2, 3, 8, 9, 10, 12, and 13 is observed.
Clonality
Polyclonal
Host
Goat
Isotype
IgG
Scientific Data Images for Human Carbonic Anhydrase IV/CA4 Antibody
Detection of Human Carbonic Anhydrase IV/CA4 by Western Blot.
Western blot shows lysates of human brain (cerebellum) tissue, human lung tisssue, and A549 human lung carcinoma cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human Carbonic Anhydrase IV/CA4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2186) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF017). A specific band was detected for Carbonic Anhydrase IV/CA4 at approximately 33 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.Detection of Human Carbonic Anhydrase IV/CA4 by Simple WesternTM.
Simple Western lane view shows lysates of human lung tissue and human kidney tissue, loaded at 0.2 mg/mL. A specific band was detected for Carbonic Anhydrase IV/CA4 at approximately 40 kDa (as indicated) using 10 µg/mL of Goat Anti-Human Carbonic Anhydrase IV/CA4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2186) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.Detection of Carbonic Anhydrase IV/CA4 in human colon.
Carbonic Anhydrase IV/CA4 was detected in immersion fixed paraffin-embedded sections of human colon using Goat Anti-Human Carbonic Anhydrase IV/CA4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2186) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to membranes of the epithelial cells in mucosal glands. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.Applications for Human Carbonic Anhydrase IV/CA4 Antibody
Application
Recommended Usage
Immunohistochemistry
3-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human colon
Sample: Immersion fixed paraffin-embedded sections of human colon
Immunoprecipitation
25 µg/mL
Sample: Conditioned cell culture medium spiked with Recombinant Human Carbonic Anhydrase IV (Catalog # 2186-CA), see our available Western blot detection antibodies
Sample: Conditioned cell culture medium spiked with Recombinant Human Carbonic Anhydrase IV (Catalog # 2186-CA), see our available Western blot detection antibodies
Simple Western
10 µg/mL
Sample: Human lung tissue and human kidney tissue
Sample: Human lung tissue and human kidney tissue
Western Blot
1 µg/mL
Sample: Human brain (cerebellum) tissue, human lung tisssue, and A549 human lung carcinoma cell line
Sample: Human brain (cerebellum) tissue, human lung tisssue, and A549 human lung carcinoma cell line
Formulation, Preparation, and Storage
Purification
Antigen Affinity-purified
Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
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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.
Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
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.
Calculators
Background: Carbonic Anhydrase IV/CA4
References
- Hewett-Emmett, D. and R.E. Tashian (1996) Mol. Phylogenet. Evol. 5:50.
- Chandrashekar, J. et al. (2009) Science 326:443.
- Rebello, G. et al. (2004) Proc. Natl. Acad. Sci. USA 101:6617.
Alternate Names
CA4, RP17
Gene Symbol
CA4
UniProt
Additional Carbonic Anhydrase IV/CA4 Products
Product Documents for Human Carbonic Anhydrase IV/CA4 Antibody
Certificate of Analysis
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Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human Carbonic Anhydrase IV/CA4 Antibody
For research use only
Citations for Human Carbonic Anhydrase IV/CA4 Antibody
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Antigen Retrieval Protocol (PIER)
- Antigen Retrieval for Frozen Sections Protocol
- Appropriate Fixation of IHC/ICC Samples
- Cellular Response to Hypoxia Protocols
- Chromogenic IHC Staining of Formalin-Fixed Paraffin-Embedded (FFPE) Tissue Protocol
- Chromogenic Immunohistochemistry Staining of Frozen Tissue
- ClariTSA™ Fluorophore Kits
- Detection & Visualization of Antibody Binding
- Fluorescent IHC Staining of Frozen Tissue Protocol
- Graphic Protocol for Heat-induced Epitope Retrieval
- Graphic Protocol for the Preparation and Fluorescent IHC Staining of Frozen Tissue Sections
- Graphic Protocol for the Preparation and Fluorescent IHC Staining of Paraffin-embedded Tissue Sections
- Graphic Protocol for the Preparation of Gelatin-coated Slides for Histological Tissue Sections
- IHC Sample Preparation (Frozen sections vs Paraffin)
- Immunofluorescent IHC Staining of Formalin-Fixed Paraffin-Embedded (FFPE) Tissue Protocol
- Immunohistochemistry (IHC) and Immunocytochemistry (ICC) Protocols
- Immunohistochemistry Frozen Troubleshooting
- Immunohistochemistry Paraffin Troubleshooting
- Immunoprecipitation Protocol
- Preparing Samples for IHC/ICC Experiments
- Preventing Non-Specific Staining (Non-Specific Binding)
- Primary Antibody Selection & Optimization
- Protocol for Heat-Induced Epitope Retrieval (HIER)
- Protocol for Making a 4% Formaldehyde Solution in PBS
- Protocol for VisUCyte™ HRP Polymer Detection Reagent
- Protocol for the Preparation & Fixation of Cells on Coverslips
- Protocol for the Preparation and Chromogenic IHC Staining of Frozen Tissue Sections
- Protocol for the Preparation and Chromogenic IHC Staining of Frozen Tissue Sections - Graphic
- Protocol for the Preparation and Chromogenic IHC Staining of Paraffin-embedded Tissue Sections
- Protocol for the Preparation and Chromogenic IHC Staining of Paraffin-embedded Tissue Sections - Graphic
- Protocol for the Preparation and Fluorescent IHC Staining of Frozen Tissue Sections
- Protocol for the Preparation and Fluorescent IHC Staining of Paraffin-embedded Tissue Sections
- Protocol for the Preparation of Gelatin-coated Slides for Histological Tissue Sections
- R&D Systems Quality Control Western Blot Protocol
- TUNEL and Active Caspase-3 Detection by IHC/ICC Protocol
- The Importance of IHC/ICC Controls
- Troubleshooting Guide: Immunohistochemistry
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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