Overview

  • 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

    Suitable for: IHC-P, WBmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Chimpanzee, Macaque monkey, Orangutan
  • 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 as ab156179)

  • 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

Applications

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.

Application 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 Use a concentration of 1 µg/ml. Detects a band of approximately 55 kDa (predicted molecular weight: 49 kDa).

Target

  • 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
    modifications

    Asn-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
  • Database links

  • Alternative names

    • CA-IX antibody
    • CA9 antibody
    • CAH9_HUMAN antibody
    • CAIX antibody
    • Carbonate dehydratase IX antibody
    • Carbonic anhydrase 9 antibody
    • Carbonic anhydrase IX antibody
    • Carbonic dehydratase antibody
    • G250 antibody
    • Membrane antigen MN antibody
    • MN antibody
    • P54/58N antibody
    • pMW1 antibody
    • RCC associated protein G250 antibody
    • RCC-associated antigen G250 antibody
    • Renal cell carcinoma-associated antigen G250 antibody
    see all

Images

  • 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.

  • 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.

References

This product has been referenced in:

See 1 Publication for this product

Customer reviews and Q&As

Application
Western blot
Loading amount
30000 cells
Gel Running Conditions
Reduced Denaturing (11)
Sample
Human Cell lysate - whole cell (MCF-7 cells (human breast adenocarcinoma cell line)
Specification
MCF-7 cells (human breast adenocarcinoma cell line
Treatment
0.5% oxygen for 16hrs
Blocking step
Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5%

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Verified customer

Submitted Dec 02 2013

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