Overview

  • Product name
    Anti-Carbonic Anhydrase IX antibody
    See all Carbonic Anhydrase IX primary antibodies
  • Description
    Rabbit polyclonal to Carbonic Anhydrase IX
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-P, Flow Cyt, IHC-Fr, WBmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide corresponding to Human Carbonic Anhydrase IX aa 359-459.
    Sequence:

    LSDTLWGPGDSRLQLNFRATQPLNGRVIEASFPAGVDSSPRAAEPVQLNS CLAAGDILALVFGLLFAVTSVAFLVQMRRQHRRGTKGGVSYRPAEVAETG A


    Database link: Q16790
    (Peptide available as ab109840)

  • Positive control
    • IHC-Fr: Human colorectal tumor tissue. IHC-P: Canine gastric mucosa tissue. WB: Ramos (human Burkitt's lymphoma cell line) Whole Cell Lysate and A549 (human lung adenocarcinoma epithelial cell line) Whole Cell Lysate

Properties

Applications

Our Abpromise guarantee covers the use of ab15086 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 1/1000 - 1/2000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. Citrate Buffer (pH 6.0)
Flow Cyt Use at an assay dependent concentration. PubMed: 25269858

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-Fr Use at an assay dependent concentration. PubMed: 20515941Slides were fixed in acetone for 20 minutes.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 55 kDa (predicted molecular weight: 49.7 kDa).Can be blocked with Recombinant Human RND1 protein (ab109840).

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

  • Formaldehyde fixed, PBS/ 0.3% Triton X-100 permeabilized frozen human colorectal tumor tissue stained for Carbonic Anhydrase IX using ab15086 (green) at a 1/1000 dilution. 5% serum used for blocking for 1 hour at RT. Primary incubation time 3 hours at RT in PBS/ 0.3% Triton X-100/ 1% BSA. Secondary: Goat polyclonal conjugated to AlexaFluor® 488 used at a 1/2000 dilution.

    See Abreview

  • All lanes : Anti-Carbonic Anhydrase IX antibody (ab15086) 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 polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Predicted band size: 49.7 kDa
    Observed band size: 55 kDa
    why is the actual band size different from the predicted?

  • ab15086 at 1/500 staining dog gastric mucosa tissue sections by IHC-P. The tissue was formaldehyde fixed and a standard citrate buffer was used for anitgen retrieval. The tissue was blocked with 10% serum prior to incubation with the antibody for 15 hours. A biotinylated donkey anti-rabbit IgG (H + L) was used as the secondary.

    See Abreview

References

This product has been referenced in:
  • Finkelmeier F  et al. Circulating hypoxia marker carbonic anhydrase IX (CA9) in patients with hepatocellular carcinoma and patients with cirrhosis. PLoS One 13:e0200855 (2018). Read more (PubMed: 30011326) »
  • Zandberga E  et al. Depletion of carbonic anhydrase IX abrogates hypoxia-induced overexpression of stanniocalcin-1 in triple negative breast cancer cells. Cancer Biol Ther 18:596-605 (2017). Read more (PubMed: 28665755) »
See all 69 Publications for this product

Customer reviews and Q&As

1-10 of 12 Abreviews or Q&A

Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa cells RIPA lysate)
Gel Running Conditions
Reduced Denaturing (4-12% precast Bis-Tris gel)
Loading amount
25 µg
Specification
HeLa cells RIPA lysate
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted May 11 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (liver)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: dako retrieval buffer
Permeabilization
No
Specification
liver
Blocking step
BSA as blocking agent for 5 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: RT°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Jan 02 2018

Application
Immunohistochemistry (Frozen sections)
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Sample
Human Tissue sections (Colorectal tumors)
Specification
Colorectal tumors
Permeabilization
Yes - 1xPBS+0.3%t-X-100
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Oct 09 2014

Question
Answer

Thank you for contacting us.

We have several HIF antibodies in our catalog that react with mouse. For example, ab8366: HIF1 alpha, ab199: HIF2 alpha, ab10134: HIF3 alpha, ab465: HIF1 beta. If you could let me know which HIF you are looking for and what application it would be used for, I would be happy to help you narrow down the results.

While we do not have any antibodies to Carbonic Anhydrase IX that are known to react with mouse, the antibodies ab108351 and ab15086 are likely to show some cross-reactivity due to sequence homology. If you would like to test either of these antibodies for use in mouse, I would be happy to offer you our 100% testing discount program. If you purchase the antibody for full price and submit an Abreview with your mouse results, you will receive a discount code that can be redeemed for a free primary antibody on a future order.

I hope this helps, please let me know if you need any additional information or assistance.

Read More

Answer

Thank you for contacting us. Unfortunately we do not have any kit corresponding to the detection of the CAIX protein. However, I would suggest to develop an ELISA assay using the CAIX protein ab114200 that would be coated on the plate. A control anti-CAIX, such as ab15086, for which the concentration is known, may be used to create a standard curve and to quantify the amount of anti-CAIX antibodies contained in the mouse sera. ab15086 is a rabbit antibody. A mouse antibody would ideal, unfortunately, the only mouse anti-CAIX we have (ab107257) is not purified. It is an ascites surpernatant and the concentration has therefore not been determined. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Answer

Thank you for your patient. Unfortunately there is no number that I can provide to you as it is impossible to know for sure how many slides you will be able to stain with a vial. This will depend on the experimental conditions, the samples and how highly expressed the protein is, and what dilution you end up choosing. This antibody is recommended to be used at a 1:1000 dilution for IHC staining as this has worked well for our lab. I would recommend ordering a single vial and test it to determine what amount will be best for your assay rather than order too much or too little without first working with the antibody. The price for a single vial is €370.00 (without shipping costs). For more information about prices and shipping conditions, please contact our Customer Service department at: orders@abcam.com Regarding your second question, we haven’t tested this system in our lab, so I cannot assure whether it will work. However, it has a fairly normal range pH in the buffer (7-8) so most likely the functions best around that range and not at a high pH. I hope this information is helpful. Please do not hesitate to contact us if you need further advice or information.

Read More

Answer

Thank you for contacting us. The concentration which is recommended for performing IHC-P with Anti-Carbonic Anhydrase IX antibody (ab15086) is 1/1000-1/2000. Taking the more concentrated range, I have estimated that from the 100µl provided (and incubating with using 200 µl of diluted antibody per slide) a total of 500 slides could be stained. For 150 slides this would equate to a total of 30µl of the vial used. This is only an estimate and optimization with the individual tissue would need to be done. In answer to your second question, although we have not tested our antibodies using the EnVision Flex+ visualization System, from having reviewed the product I seen no reason why it would not be compatible. I hope this information has been of help. If you have any further questions please do not hesitate to contact us again.

Read More

Answer

Thank you for your reply. It seems from your description that you are doing everything fine. If I understand correctly, you are using the peptide at 1 ug/mL and the antibody at 0.1 ug/mL (1/10,000 dilution). At same volumes, the peptide is being used at 10-fold excess. Since this gave only a small effect in blocking, I would suggest trying the peptide at a 100-fold excess. As for the incubation time: if overnight incubation at 4 degree worked a bit better, I would stick with this. Should the increase in peptide not help, please let me know. If the peptide was purchased within the last 6 months, we can discuss options of replacement, credit or refund. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Dog Tissue sections (Soft Tissue Sarcoma, Gastric Mucosa)
Specification
Soft Tissue Sarcoma, Gastric Mucosa
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10%

Abcam user community

Verified customer

Submitted Dec 13 2006

Answer

Thank you for your enquiry. I enquired with our source for this antibody and there have not been any reports of this problem where the antibody is staining muscle. Ab15086 was characterized for use in IHC with renal carcinoma tissue and I would recommend that as a positive control. If you can provide some additional details regarding the protocol that was used, we can further investigate this issue. An image would be helpful as well.

Read More

1-10 of 12 Abreviews or Q&A

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