Recombinant Anti-Carbonic Anhydrase 12/CA12 antibody [EPR14861] - C-terminal (ab195233)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR14861] to Carbonic Anhydrase 12/CA12 - C-terminal
- Suitable for: Flow Cyt (Intra), IHC-P, WB
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-Carbonic Anhydrase 12/CA12 antibody [EPR14861] - C-terminal
See all Carbonic Anhydrase 12/CA12 primary antibodies -
Description
Rabbit monoclonal [EPR14861] to Carbonic Anhydrase 12/CA12 - C-terminal -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), IHC-P, WBmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- MCF7, SKOV3, 293, HeLa, A549, Human fetal kidney, Human ovary cancer and Human breast cancer lysates; Human renal adenocarcinoma, ovarian carcinoma, bladder transitional cell carcinoma, pancreas, colon and stomach tissues; SK-OV-3 cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), PBS, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR14861 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Conjugation kits
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab195233 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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Flow Cyt (Intra) |
1/130.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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IHC-P | (1) |
1/50 - 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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WB |
1/10000. Detects a band of approximately 45 kDa (predicted molecular weight: 39 kDa).
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Notes |
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Flow Cyt (Intra)
1/130. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
1/10000. Detects a band of approximately 45 kDa (predicted molecular weight: 39 kDa). |
Target
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Function
Reversible hydration of carbon dioxide. -
Tissue specificity
Highly expressed in colon, kidney, prostate, intestine and activated lymphocytes. Expressed at much higher levels in the renal cell cancers than in surrounding normal kidney tissue. Moderately expressed in pancreas, ovary and testis. -
Involvement in disease
Defects in CA12 are the cause of hyperchlorhidrosis isolated (HCHLH) [MIM:143860]. HCHLH is a disorder characterized by excessive sweating and increased sweat chloride levels. Affected individuals suffer from episodes of hyponatremic dehydration and report increased amounts of visible salt precipitates in sweat. -
Sequence similarities
Belongs to the alpha-carbonic anhydrase family. -
Cellular localization
Membrane. - Information by UniProt
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Database links
- Entrez Gene: 771 Human
- Omim: 603263 Human
- SwissProt: O43570 Human
- Unigene: 210995 Human
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Alternative names
- CA 12 antibody
- CA XII antibody
- CA-XII antibody
see all
Images
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All lanes : Anti-Carbonic Anhydrase 12/CA12 antibody [EPR14861] - C-terminal (ab195233) at 1/10000 dilution
Lane 1 : Wild-type A549 (Human lung carcinoma cell line) whole cell lysate
Lane 2 : Ca12 knockout A549 (Human lung carcinoma cell line) whole cell lysate
Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 39 kDa
Observed band size: 40 kDa why is the actual band size different from the predicted?Lanes 1 - 3: Merged signal (red and green). Green - ab195233 observed at 40 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
ab195233 was shown to react with Carbonic Anhydrase 12/CA12 in wild-type A549 cells in Western blot with loss of signal observed in Ca12 knockout cell line ab273738 (knockout cell lysate ab273778). Wild-type A549 and Ca12 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab195233 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
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All lanes : Anti-Carbonic Anhydrase 12/CA12 antibody [EPR14861] - C-terminal (ab195233) at 1/10000 dilution
Lane 1 : MCF7 cell lysate
Lanes 2 & 5 : SKOV3 cell lysate
Lane 3 : 293 cell lysate
Lane 4 : HeLa cell lysate
Lane 6 : Human fetal kidney lysate
Lane 7 : Human ovary cancer lysate
Lane 8 : Human breast cancer lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution
Developed using the ECL technique.
Predicted band size: 39 kDa
Exposure time: 10 seconds -
Intracellular flow cytometric analysis of SK-OV-3cells (paraformaldehyde-fixed, 2%) labeling CA12 with ab195233 at 1/130 dilution (red) or a Rabbit monoclonal IgG (negative) (black), followed by Goat anti rabbit IgG (FITC) secondary at 1/150 dilution. Unlabeled cells (Blue).
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Immunohistochemical analysis of paraffin-embedded Human stomach tissue labeling CA12 with ab195233 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution and counter-stained with Hematoxylin. (inset: negative control).
Note: Cell membrane and cytoplasm staining on human stomach tissue was observed.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded human colon tissue labeling CA12 with ab195233 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution and counter-stained with Hematoxylin. (inset: negative control).
Note: Cell membrane and cytoplasm staining on human colon tissue was observed.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling CA12 with ab195233 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution and counter-stained with Hematoxylin. (inset: negative control).
Note: Cell membrane and weakly cytoplasm staining on human pancreas tissue was observed.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded Human bladder transitional cell carcinoma tissue labeling CA12 with ab195233 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution and counter-stained with Hematoxylin. (inset: negative control).
Note: Cell membrane and weakly cytoplasm staining on human transitional cell carcinoma of bladder tissue was observed.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded Human ovarian carcinoma tissue labeling CA12 with ab195233 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution and counter-stained with Hematoxylin. (inset: negative control).
Note: Cell membrane and weakly cytoplasm staining on human ovarian carcinoma tissue was observed.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded Human renal adenocarcinoma tissue labeling CA12 with ab195233 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution and counter-stained with Hematoxylin. (inset: negative control).
Note: Cell membrane and cytoplasm staining on human renal adenocarcinoma tissue was observed.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (5)
ab195233 has been referenced in 5 publications.
- Huang S et al. In vivo development and single-cell transcriptome profiling of human brain organoids. Cell Prolif 55:e13201 (2022). PubMed: 35141969
- Tavares-Valente D et al. Disruption of pH Dynamics Suppresses Proliferation and Potentiates Doxorubicin Cytotoxicity in Breast Cancer Cells. Pharmaceutics 13:N/A (2021). PubMed: 33572458
- Zhang X et al. Endogenous glutamate determines ferroptosis sensitivity via ADCY10-dependent YAP suppression in lung adenocarcinoma. Theranostics 11:5650-5674 (2021). PubMed: 33897873
- Chafe SC et al. Genome-wide synthetic lethal screen unveils novel CAIX-NFS1/xCT axis as a targetable vulnerability in hypoxic solid tumors. Sci Adv 7:N/A (2021). PubMed: 34452919
- Huang T et al. Carbonic anhydrase 12 gene silencing reverses the sensitivity of paclitaxel in drug-resistant breast cancer cells. Bioengineered 12:9806-9818 (2021). PubMed: 34696661