Anti-Carbonic Anhydrase 9/CA9 antibody (ab128883)
Key features and details
- Rabbit polyclonal to Carbonic Anhydrase 9/CA9
- Suitable for: IHC-P, WB
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-Carbonic Anhydrase 9/CA9 antibody
See all Carbonic Anhydrase 9/CA9 primary antibodies -
Description
Rabbit polyclonal to Carbonic Anhydrase 9/CA9 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species IHC-P HumanWB Human
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Immunogen
Synthetic peptide corresponding to Human Carbonic Anhydrase 9/CA9 aa 400 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
(Peptide available asab156179) -
Positive control
- This antibody gave a positive signal in both A549 and Ramos whole cell lysates within WB as well as in Human kidney carcinoma formalin fixed paraffin embedded tissue section within IHC.
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 or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab128883 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
GuaranteedTested applications are guaranteed to work and covered by our Abpromise guarantee.
PredictedPredicted to work for this combination of applications and species but not guaranteed.
IncompatibleDoes not work for this combination of applications and species.
Application Species IHC-P HumanWB HumanAll applications ChimpanzeeMacaque monkeyOrangutanApplication Abreviews Notes IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.WB (1) Use a concentration of 1 µg/ml. Detects a band of approximately 55 kDa (predicted molecular weight: 49 kDa).Notes IHC-P
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.WB
Use a concentration of 1 µg/ml. Detects a band of approximately 55 kDa (predicted molecular weight: 49 kDa).Target
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Function
Reversible hydration of carbon dioxide. Participates in pH regulation. May be involved in the control of cell proliferation and transformation. Appears to be a novel specific biomarker for a cervical neoplasia. -
Tissue specificity
Expressed primarily in carcinoma cells lines. Expression is restricted to very few normal tissues and the most abundant expression is found in the epithelial cells of gastric mucosa. -
Sequence similarities
Belongs to the alpha-carbonic anhydrase family.
Contains 1 alpha-carbonic anhydrase domain. -
Post-translational
modificationsAsn-346 bears high-mannose type glycan structures. -
Cellular localization
Nucleus. Nucleus, nucleolus. Cell membrane. Cell projection, microvillus membrane. Found on the surface microvilli and in the nucleus, particularly in nucleolus. - Information by UniProt
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Database links
- Entrez Gene: 768 Human
- Omim: 603179 Human
- SwissProt: Q16790 Human
- Unigene: 63287 Human
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Alternative names
- CA-IX antibody
- CA9 antibody
- CAH9_HUMAN antibody
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Carbonic Anhydrase 9/CA9 antibody (ab128883)
IHC image of Carbonic Anhydrase 9/CA9 staining in Human kidney carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab128882, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
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Western blot - Anti-Carbonic Anhydrase 9/CA9 antibody (ab128883)All lanes : Anti-Carbonic Anhydrase 9/CA9 antibody (ab128883) at 1 µg/ml
Lane 1 : Ramos (Human Burkitt's lymphoma cell line) Whole Cell Lysate
Lane 2 : A549 (Human lung adenocarcinoma epithelial cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 49 kDa
Observed band size: 55 kDa why is the actual band size different from the predicted?
Additional bands at: 36 kDa, 50 kDa, 85 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 12 minutes
Carbonic Anhydrase IX contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted. This blot was produced using a 10% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab128883 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.
Protocols
Datasheets and documents
References (2)
ab128883 has been referenced in 2 publications.
- Frohwitter G et al. Site-specific gene expression patterns in oral cancer. Head Face Med 13:6 (2017). IHC-P ; Human . PubMed: 28486965
- Frohwitter G et al. Cytokeratin and protein expression patterns in squamous cell carcinoma of the oral cavity provide evidence for two distinct pathogenetic pathways. Oncol Lett 12:107-113 (2016). PubMed: 27347109
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Carbonic Anhydrase 9/CA9 antibody (ab128883)
IHC image of Carbonic Anhydrase 9/CA9 staining in Human kidney carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab128882, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
-
Western blot - Anti-Carbonic Anhydrase 9/CA9 antibody (ab128883)All lanes : Anti-Carbonic Anhydrase 9/CA9 antibody (ab128883) at 1 µg/ml
Lane 1 : Ramos (Human Burkitt's lymphoma cell line) Whole Cell Lysate
Lane 2 : A549 (Human lung adenocarcinoma epithelial cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 49 kDa
Observed band size: 55 kDa why is the actual band size different from the predicted?
Additional bands at: 36 kDa, 50 kDa, 85 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 12 minutes
Carbonic Anhydrase IX contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted. This blot was produced using a 10% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab128883 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.
