Anti-Carbonic Anhydrase IX antibody (ab10471)

Overview

  • Product nameAnti-Carbonic Anhydrase IX antibodySee all Carbonic Anhydrase IX primary antibodies ...
  • Description
    Rabbit polyclonal to Carbonic Anhydrase IX
  • Tested applicationsWB, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 100 - 200 of Human Carbonic Anhydrase IX.

    (Peptide available as ab47011.)

  • Positive control
    • Carbonic Anhydrase IX recombinant protein This antibody gave a positive result in IF in the following methanol fixed cell lines: DU145.

Properties

Applications

Our Abpromise guarantee covers the use of ab10471 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
WB Use at an assay dependent dilution. Predicted molecular weight: 49.7 kDa.Can be blocked with Human Carbonic Anhydrase IX peptide (ab47011).
IHC-P 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF Use a concentration of 5 µg/ml.

Target

  • FunctionReversible 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 specificityExpressed 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 similaritiesBelongs to the alpha-carbonic anhydrase family.
  • Post-translational
    modifications
    Asn-346 bears high-mannose type glycan structures.
  • Cellular localizationNucleus. 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
    • 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
    • P54/58N antibody
    • pMW1 antibody
    • RCC associated protein G250 antibody
    • RCC-associated antigen G250 antibody
    • Renal cell carcinoma associated antigen G250 antibody
    • Renal cell carcinoma-associated antigen G250 antibody
    see all

Anti-Carbonic Anhydrase IX antibody images

  • Anti-Carbonic Anhydrase IX antibody (ab10471) at 1 µg/ml + Carbonic Anhydrase IX recombinant protein at 0.1 µg/ml

    Secondary
    Goat polyclonal to Rabbit IgG H&L (HRP) Pre-Adsorbed at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 49.7 kDa
    Observed band size : 53,54 kDa (why is the actual band size different from the predicted?)
  • ab10471 staining Carbonic Anhydrase IX in Human kidney tissue sections by IHC-P (Formaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 5 minutes of peroxidase block followed by 10 minutes of protein block at 20°C; antigen retrieval was heat mediated in retrieval solution. Samples were incubated with primary antibody (1/250 in antibody diluent) for 45 minutes at 20°C. An undiluted HRP-conjugated polymer goat anti-mouse/rabbit IgG polyclonal was used as secondary antibody.

    See Abreview

  • ICC/IF image of ab10471 stained DU145 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab10471 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References for Anti-Carbonic Anhydrase IX antibody (ab10471)

ab10471 has not yet been referenced specifically in any publications.

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (kidney)
Specification kidney
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Dako Target Retreival Solution
Blocking step 5 minutes of peroxidase block then 10 minutes of protein block. These are ready-to-use reagents purchased from Dako as blocking agent for 15 minute(s) · Concentration: 100% · Temperature: 20°C
Username

Mr. Antibody Solutions

Verified customer

Submitted Mar 18 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (MCF-7, 786-O, RCC4)
Loading amount 20 µg
Specification MCF-7, 786-O, RCC4
Treatment MCF-7: 21% oxygen vs 1% oxygen for 20 hours
Gel Running Conditions Reduced Denaturing (Pierce precise protein gel 4-20%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted Aug 07 2009

Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - whole (Cortical lysates)
Loading amount 20 µg
Specification Cortical lysates
Gel Running Conditions Reduced Denaturing (10% gel)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Dr. Martin Broadstock

Verified customer

Submitted May 13 2009

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"