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    carbonic-anhydrase-9ca9-antibody-ab15086.pdf

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Cardiovascular Hypoxia Associated Proteins
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Anti-Carbonic Anhydrase 9/CA9 antibody (ab15086)

  • Datasheet
  • SDS
Reviews (4)Q&A (8)References (123)

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Western blot - Anti-Carbonic Anhydrase 9/CA9 antibody (ab15086)
  • Western blot - Anti-Carbonic Anhydrase 9/CA9 antibody (ab15086)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Carbonic Anhydrase 9/CA9 antibody (ab15086)
  • Immunohistochemistry (Frozen sections) - Anti-Carbonic Anhydrase 9/CA9 antibody (ab15086)
  • Western blot - Anti-Carbonic Anhydrase 9/CA9 antibody (ab15086)

Key features and details

  • Rabbit polyclonal to Carbonic Anhydrase 9/CA9
  • Suitable for: Flow Cyt (Intra), ICC, IHC-Fr, IHC-P, WB
  • Reacts with: Rat, Human
  • Isotype: IgG

Get better batch-to-batch reproducibility with a recombinant antibody

Product image
Anti-Carbonic Anhydrase 9/CA9 antibody [EPR4151(2)] (ab108351)
  • Research with confidence – consistent and reproducible results with every batch
  • Long-term and scalable supply – powered by recombinant technology for fast production
  • Success from the first experiment – confirmed specificity through extensive validation
  • Ethical standards compliant – production is animal-free

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: Flow Cyt (Intra), ICC, IHC-Fr, IHC-P, WBmore details
  • Species reactivity

    Reacts with: Rat, Human
  • Immunogen

    Synthetic peptide corresponding to Human Carbonic Anhydrase 9/CA9 aa 359-459.
    Sequence:

    LSDTLWGPGDSRLQLNFRATQPLNGRVIEASFPAGVDSSPRAAEPVQLNS CLAAGDILALVFGLLFAVTSVAFLVQMRRQHRRGTKGGVSYRPAEVAETG A


    Database link: Q16790
    (Peptide available as ab109840)
    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • Positive control

    • WB: Ramos, A549, HeLa and MDA-MB-231 cell lysates; Rat renal cortex lysate. Intr Flow Cyt: A-431 and U-87 MG cells. ICC: A-431 cells. IHC-Fr: Human renal cell cancer tumor sections. IHC-P: Human breast cancer sections.
  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.40
    Preservative: 0.1% Sodium azide
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Cardiovascular
    • Hypoxia
    • Associated Proteins
    • Signal Transduction
    • Metabolism
    • Vitamins / Minerals
    • Cancer
    • Cancer Metabolism
    • Response to hypoxia
    • Metabolism
    • Pathways and Processes
    • Cofactors, Vitamins / minerals
    • Vitamins / minerals
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Hypoxia
    • Metabolism
    • Types of disease
    • Cancer

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Immunizing Peptide (Blocking)

    • Recombinant Human RND1 protein (ab109840)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)

Applications

The Abpromise guarantee

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
Flow Cyt (Intra)
1/1000.
ICC
Use a concentration of 2 - 5 µg/ml.
IHC-Fr (1)
1/200 - 1/500.
IHC-P (2)
1/200 - 1/500.
WB (1)
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).
Notes
Flow Cyt (Intra)
1/1000.
ICC
Use a concentration of 2 - 5 µg/ml.
IHC-Fr
1/200 - 1/500.
IHC-P
1/200 - 1/500.
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.
  • Target information above from: UniProt accession Q16790 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 768 Human
    • Entrez Gene: 313495 Rat
    • Omim: 603179 Human
    • SwissProt: Q16790 Human
    • Unigene: 63287 Human
    • 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

    • Western blot - Anti-Carbonic Anhydrase 9/CA9 antibody (ab15086)
      Western blot - Anti-Carbonic Anhydrase 9/CA9 antibody (ab15086)
      Western blot analysis labeling Carbonic Anhydrase 9/CA9 with ab15086 at 1 ug/ml in rat renal cortex lysate.
    • Western blot - Anti-Carbonic Anhydrase 9/CA9 antibody (ab15086)
      Western blot - Anti-Carbonic Anhydrase 9/CA9 antibody (ab15086)
      Western blot analysis labeling Carbonic Anhydrase 9/CA9 with ab15086 at 1 ug/ml in 1) HeLa, 2) MDA-MB-231, and 3) A549 whole cell lysates.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Carbonic Anhydrase 9/CA9 antibody (ab15086)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Carbonic Anhydrase 9/CA9 antibody (ab15086)
      Immunohistochemistry analysis of a formalin-fixed, paraffin-embedded tissue section of human breast cancer using ab15086 at 1:1000 dilution. The primary antibody bound to Carbonic Anydrase 9/CA9 antigens in the tissue section was detected using a HRP labeled secondary antibody and DAB reagent. Nuclei of the cells were counterstained with hematoxylin. This antibody generated an expected cytoplasmic staining of Carbonic Anydrase 9/CA9 protein with an intense signal around the cellular membranes in tumor cores. The latter are more likely to be hypoxic in growing tumors which signifies that the observed Carbonic Anydrase 9/CA9 staining is specific.
    • Immunohistochemistry (Frozen sections) - Anti-Carbonic Anhydrase 9/CA9 antibody (ab15086)
      Immunohistochemistry (Frozen sections) - Anti-Carbonic Anhydrase 9/CA9 antibody (ab15086)
      Immunohistochemistry analysis of human renal cell cancer tumor cryosections stained with ab15086 at a 1/500 dilution (left) or normal rabbit serum (right).
    • Western blot - Anti-Carbonic Anhydrase 9/CA9 antibody (ab15086)
      Western blot - Anti-Carbonic Anhydrase 9/CA9 antibody (ab15086)
      All lanes : Anti-Carbonic Anhydrase 9/CA9 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?

    Protocols

    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (123)

    Publishing research using ab15086? Please let us know so that we can cite the reference in this datasheet.

    ab15086 has been referenced in 123 publications.

    • Krawczyk MA  et al. High Expression of Solute Carrier Family 2 Member 1 (SLC2A1) in Cancer Cells Is an Independent Unfavorable Prognostic Factor in Pediatric Malignant Peripheral Nerve Sheath Tumor. Diagnostics (Basel) 11:N/A (2021). PubMed: 33810575
    • Gournay M  et al. Renal cell carcinoma with leiomyomatous stroma in tuberous sclerosis complex: a distinct entity. Virchows Arch 478:793-799 (2021). PubMed: 32845354
    • Le Joncour V  et al. Targeting the Urotensin II/UT G Protein-Coupled Receptor to Counteract Angiogenesis and Mesenchymal Hypoxia/Necrosis in Glioblastoma. Front Cell Dev Biol 9:652544 (2021). PubMed: 33937253
    • Schneider MA  et al. Phase Ib dose-escalation study of the hypoxia-modifier Myo-inositol trispyrophosphate in patients with hepatopancreatobiliary tumors. Nat Commun 12:3807 (2021). PubMed: 34155211
    • Kallio P  et al. Blocking Angiopoietin-2 Promotes Vascular Damage and Growth Inhibition in Mouse Tumors Treated with Small Doses of Radiation. Cancer Res 80:2639-2650 (2020). PubMed: 32312835
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 12 Abreviews or Q&A

    Western blot abreview for Anti-Carbonic Anhydrase IX antibody

    Good
    Abreviews
    Abreviews
    abreview image
    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
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted May 11 2018

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Carbonic Anhydrase 9/CA9 antibody

    Good
    Abreviews
    Abreviews
    abreview image
    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
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Jan 02 2018

    Immunohistochemistry (Frozen sections) abreview for Anti-Carbonic Anhydrase IX antibody

    Good
    Abreviews
    Abreviews
    abreview image
    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
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Oct 09 2014

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Carbonic Anhydrase IX antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    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%
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Dec 13 2006

    Question

    looking for CA-9 mouse antibodies looking for HIF antibodies

    Read More

    Abcam community

    Verified customer

    Asked on Jun 29 2012

    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

    Abcam Scientific Support

    Answered on Jun 29 2012

    Question

    Hello, I am writing you re your Indirect ELISA kits and hope you can advise which kit buy and protocol to use. We have developed RCC mouse model using  RENCA mouse RCC cells over expressing the human CAIX protein. We would like to test/control for the possible presence of mouse anti human CAIX antibodies in the mouse sera. That is to say we would like to coat the plates with recombinant human CAIX protein, use OUR mice sera as primary Abs (as a “detector Antibodies”) and then follow with a secondary Anti Mouse Ab  HRP Conjugated/other detection conjugate. Could you possibly advise which kit buy and protocol to use. Thank you

    Read More

    Abcam community

    Verified customer

    Asked on Dec 05 2011

    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

    Abcam Scientific Support

    Answered on Dec 05 2011

    Question

    Dear Colleques, I would like to order antibody against CA IX (ab 15086). I need to use them for 150 formalin fixed, paraffin embeded sections. Let me know about the amount I need to order, wheter they work with ENVISION FLEX+, HIGH pH, FOR AS, and about the price in total. I need these information as soon as possible. Best regards,

    Read More

    Abcam community

    Verified customer

    Asked on Nov 30 2011

    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

    Abcam Scientific Support

    Answered on Nov 30 2011

    Question

    Greetings-   I would like to kindly ask you for any information on the following topic: my customer would like to purchase the ab15086 for IHC-P and he is asking what is the efficiency of this antibody for 1.5 mmx15mm fragment? In other words, how much this antibody he has to use to prepare 150 sections/fragments ? Can he work on EnVision Flex+ visualization System with abcam antibodies?   Thank you in advance for your help, I really appreciate it.   Best regards,    

    Read More

    Abcam community

    Verified customer

    Asked on Nov 30 2011

    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

    Abcam Scientific Support

    Answered on Nov 30 2011

    Question

    I have only tried changing the preincubation step (antibody with blocking peptide) at different amounts of time (so, once overnight at 4 degrees and once for 30 mins at room temperature). The one overnight did help a bit but the bands that should be blocked were still quite bright. To answer you questions:   1) What samples did you use (sample type, species)? I am using lysed keratinocytes infected with HPV 16 2) What antibody dilution have you tried? I have tried many antibody dilutions ranging from 1:200 to 1:10000 and I have found that 1:10000 gives me the best result (no background, but bands of interest still nice and bright). 3) What blocking reagent have you been using? The blocking reagent I use is 5% milk in 1XTBST 4) Which peptide dilutions have you tried? How long did you incubate the antibody with the peptide before the doing the Western blot?  For the peptide, I have only tried the suggested amount of 1ug/mL. As stated above, I have tried two different times; 30 minutes at room temperature or overnight at 4degrees. Please let me know if it is obvious that I have missed something here or if I should just try a higher concentration of peptide or perhaps a longer preincubation at room temperature.

    Read More

    Abcam community

    Verified customer

    Asked on Nov 08 2011

    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

    Abcam Scientific Support

    Answered on Nov 08 2011

    Question

    BATCH NUMBER -- NOT SPECIFIED -- ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM Non-specific staining SAMPLE human colorectal cancer PRIMARY ANTIBODY ab15086 carbonic anhydrase IX ANTIBODY STORAGE CONDITIONS just bought FIXATION OF SAMPLE formaldehyde ANTIGEN RETRIEVAL citrate HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 1 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes ADDITIONAL NOTES I can't answer most ofthe qustions above as I am not the individual who does the specific procedure. But the question is: Using IHC, we are seeing very high staining of muscle. We see the expected membrane staining associated with CAIX staining in a few areas of the tumor, but surmise that the antibody is cross-reacting with something else in muscle. Using identical reagents (but different primary) we do not see any of this staininig. Are you aware of anyone else who has had this problem? I see no comment about it in the literature. All thoughts appreciated.

    Read More

    Abcam community

    Verified customer

    Asked on Jan 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

    Abcam Scientific Support

    Answered on Jan 16 2006

    1-10 of 12 Abreviews or Q&A

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