Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Carbonic anhydrase 2/CA2 antibody [EPR5195] (ab124687)

Reviews (1)Q&A (2)References (12)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Carbonic anhydrase 2/CA2 antibody [EPR5195] (ab124687)
  • Western blot - Anti-Carbonic anhydrase 2/CA2 antibody [EPR5195] (ab124687)
  • Western blot - Anti-Carbonic anhydrase 2/CA2 antibody [EPR5195] (ab124687)
  • Western blot - Anti-Carbonic anhydrase 2/CA2 antibody [EPR5195] (ab124687)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Carbonic anhydrase 2/CA2 antibody [EPR5195] (ab124687)
  • OI-RD Scanning - Anti-Carbonic anhydrase 2/CA2 antibody [EPR5195] (ab124687)
  • Anti-Carbonic anhydrase 2/CA2 antibody [EPR5195] (ab124687)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR5195] to Carbonic anhydrase 2/CA2
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-Carbonic anhydrase 2/CA2 antibody [EPR5195]
    See all Carbonic anhydrase 2/CA2 primary antibodies

  • Description

    Rabbit monoclonal [EPR5195] to Carbonic anhydrase 2/CA2

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    IHC-P
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide corresponding to Human Carbonic anhydrase 2/CA2 aa 200-300 (C terminal).

  • Positive control

    • WB: A431, Caco 2 and HEK293T cell lysates; Human heart and fetal brain lysates. IHC-P: Human kidney and brain tissues.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    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 the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.

  • Dissociation constant (KD)

    KD = 5.42 x 10 -10 M
    Learn more about KD

  • Storage buffer

    pH: 7.2
    Preservative: 0.05% Sodium azide
    Constituents: 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant
  • Concentration information loading...

  • Purity

    Tissue culture supernatant

  • Clonality

    Monoclonal

  • Clone number

    EPR5195

  • Isotype

    IgG

  • Research areas

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab124687 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
IHC-P
Human
WB
Human
All applications
Mouse
Rat
Application Abreviews Notes
WB (1)
1/1000 - 1/10000. Detects a band of approximately 29 kDa (predicted molecular weight: 29 kDa).
IHC-P
1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Notes
WB
1/1000 - 1/10000. Detects a band of approximately 29 kDa (predicted molecular weight: 29 kDa).
IHC-P
1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
  • Application notes
    Is unsuitable for Flow Cyt,ICC/IF or IP.

    • Target

    • Function

      Essential for bone resorption and osteoclast differentiation (By similarity). Reversible hydration of carbon dioxide. Can hydrates cyanamide to urea. Involved in the regulation of fluid secretion into the anterior chamber of the eye.

    • Involvement in disease

      Defects in CA2 are the cause of osteopetrosis autosomal recessive type 3 (OPTB3) [MIM:259730]; also known as osteopetrosis with renal tubular acidosis, carbonic anhydrase II deficiency syndrome, Guibaud-Vainsel syndrome or marble brain disease. Osteopetrosis is a rare genetic disease characterized by abnormally dense bone, due to defective resorption of immature bone. The disorder occurs in two forms: a severe autosomal recessive form occurring in utero, infancy, or childhood, and a benign autosomal dominant form occurring in adolescence or adulthood. Autosomal recessive osteopetrosis is usually associated with normal or elevated amount of non-functional osteoclasts. OPTB3 is associated with renal tubular acidosis, cerebral calcification (marble brain disease) and in some cases with mental retardation.

    • Sequence similarities

      Belongs to the alpha-carbonic anhydrase family.

    • Cellular localization

      Cytoplasm.
    • Information by UniProt

    • Database links

      see all

    • Alternative names

      • CA 2 antibody
      • CA II antibody
      • CA-II antibody
      • Ca2 antibody
      • CAC antibody
      • CAH2_HUMAN antibody
      • CAII antibody
      • Car 2 antibody
      • Car2 antibody
      • Carbonate dehydratase II antibody
      • Carbonic anhydrase 2 antibody
      • Carbonic anhydrase B antibody
      • Carbonic anhydrase C antibody
      • Carbonic anhydrase C, formerly antibody
      • Carbonic anhydrase II antibody
      • Carbonic dehydratase antibody
      • epididymis luminal protein 76 antibody
      • Epididymis secretory protein Li 282 antibody
      • HEL-76 antibody
      • HEL-S-282 antibody
      see all

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Carbonic anhydrase 2/CA2 antibody [EPR5195] (ab124687)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Carbonic anhydrase 2/CA2 antibody [EPR5195] (ab124687)

      ab124687, at 1/250 dilution, staining Carbonic anhydrase 2/CA2 in paraffin-embedded Human kidney tissue by Immunohistochemistry.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Western blot - Anti-Carbonic anhydrase 2/CA2 antibody [EPR5195] (ab124687)
      Western blot - Anti-Carbonic anhydrase 2/CA2 antibody [EPR5195] (ab124687)
      All lanes : Anti-Carbonic anhydrase 2/CA2 antibody [EPR5195] (ab124687) at 1/1000 dilution

      Lane 1 : Wild-type HEK293T cell lysate
      Lane 2 : CA2 knockout HEK293T cell lysate
      Lane 3 : A431 cell lysate
      Lane 4 : Caco-2 cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

      Predicted band size: 29 kDa
      Observed band size: 29 kDa



      Lanes 1-4: Merged signal (red and green). Green - ab124687 observed at 29 kDa. Red - loading control ab8245 observed at 36 kDa. 

       ab124687 Anti-Carbonic anhydrase 2/CA2 antibody [EPR5195] was shown to specifically react with Carbonic anhydrase 2/CA2 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab265072 (knockout cell lysate ab257084) was used.  Wild-type and Carbonic anhydrase 2/CA2 knockout samples were subjected to SDS-PAGE.  ab124687 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-Carbonic anhydrase 2/CA2 antibody [EPR5195] (ab124687)
      Western blot - Anti-Carbonic anhydrase 2/CA2 antibody [EPR5195] (ab124687)

      Lane 1: Wild-type HAP1 cell lysate (40 µg)
      Lane 2: Carbonic anhydrase 2/CA2 knockout HAP1 cell lysate (40 µg)
      Lane 3: A431 cell lysate (40 µg)
      Lane 4: HEK293 cell lysate (40 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab124687 observed at 32 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab124687 was shown to specifically react with Carbonic anhydrase 2/CA2 when Carbonic anhydrase 2/CA2 knockout samples were used. Wild-type and Carbonic anhydrase 2/CA2 knockout samples were subjected to SDS-PAGE. Ab124687 and ab8245 (loading control to GAPDH) were diluted at 1/1000 and 1:10,000 dilution respectively and incubated overnight at 4C. Blots were developed with IRDye® 800CW Goat anti-Rabbit IgG (H + L) and IRDye® 680 Goat anti-Mouse IgG (H + L) secondary antibodies at 1:10,000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-Carbonic anhydrase 2/CA2 antibody [EPR5195] (ab124687)
      Western blot - Anti-Carbonic anhydrase 2/CA2 antibody [EPR5195] (ab124687)
      All lanes : Anti-Carbonic anhydrase 2/CA2 antibody [EPR5195] (ab124687) at 1/1000 dilution

      Lane 1 : Human fetal brain tissue lysate
      Lane 2 : Caco 2 cell lysate
      Lane 3 : 293T cell lysate
      Lane 4 : Human heart tissue lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat anti-rabbit HRP at 1/2000 dilution

      Predicted band size: 29 kDa
      Observed band size: 29 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Carbonic anhydrase 2/CA2 antibody [EPR5195] (ab124687)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Carbonic anhydrase 2/CA2 antibody [EPR5195] (ab124687)

      ab124687, at 1/250 dilution, staining Carbonic anhydrase 2/CA2 in paraffin-embedded Human brain tissue by Immunohistochemistry.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • OI-RD Scanning - Anti-Carbonic anhydrase 2/CA2 antibody [EPR5195] (ab124687)
      OI-RD Scanning - Anti-Carbonic anhydrase 2/CA2 antibody [EPR5195] (ab124687)
      Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD
    • Anti-Carbonic anhydrase 2/CA2 antibody [EPR5195] (ab124687)
      Anti-Carbonic anhydrase 2/CA2 antibody [EPR5195] (ab124687)

    References (12)

    Publishing research using ab124687? Please let us know so that we can cite the reference in this datasheet.

    ab124687 has been referenced in 12 publications.

    • Li S  et al. PKC-d deficiency in B cells displays osteopenia accompanied with upregulation of RANKL expression and osteoclast-osteoblast uncoupling. Cell Death Dis 11:762 (2020). PubMed: 32938907
    • Hannen R  et al. Comparative Transcriptomic Analysis of Temozolomide Resistant Primary GBM Stem-Like Cells and Recurrent GBM Identifies Up-Regulation of the Carbonic Anhydrase CA2 Gene as Resistance Factor. Cancers (Basel) 11:N/A (2019). PubMed: 31262047
    • Suri BK  et al. Carbonic anhydrases in human keratinocytes and their regulation by all-trans retinoic acid and 1a,25-dihydroxyvitamin D3. Exp Dermatol 28:976-980 (2019). PubMed: 31125456
    • Li H  et al. Expression and localization of Forkhead transcription factor A1 in the three-dimensional reconstructed eccrine sweat glands. Acta Histochem 120:520-524 (2018). PubMed: 29909922
    • Suri BK  et al. Toll-like Receptor 3 Agonist, Polyinosinic-polycytidylic Acid, Upregulates Carbonic Anhydrase II in Human Keratinocytes. Acta Derm Venereol 98:762-765 (2018). PubMed: 29738048
    View all Publications for this product

    Customer reviews and Q&As

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    1-3 of 3 Abreviews or Q&A

    Western blot abreview for Anti-Carbonic Anhydrase II antibody [EPR5195]

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    10 µg
    Gel Running Conditions
    Reduced Denaturing (12.5% gel)
    Sample
    Human Cell lysate - whole cell (293T)
    Specification
    293T
    Treatment
    CA2-3xflag transfected
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Jan 30 2015

    Question

    I'm looking into a panel of antibodies from your company to use in cells of pig origin. ab758 ab27988 ab7504 ab124687 ab28028 ab11333 ab32362 I would like to know the following: Have they been tested in pig cells? If yes, are they reactive towards pig cells? If you do not have that information, can you tell me the epitope or peptide sequence that was used in generating that antibody? If you do not want to tell me that info, can you tell me the identity percentage of match of the pig compared to the epitope that it was used in generating the antibody. Many thanks!

    Read More
    Answer

    I heard back from the lab about the last two antibodies.
    For ab32362, the immunogen sequence shares 100% homology with pig.
    For ab124687, the immunogen peptide maps between amino acids 235 - 260, on the C-terminus of human CA2 (UniProt: P00918).
    I hope this information helps. Please contact us with any other question.

    Read More

    Question


    Inquiry: I'm looking into a panel of antibodies from your company to use in cells of pig origin. ab758 ab27988 ab7504 ab124687 ab28028 ab11333 ab32362 I would like to know the following: Have they been tested in pig cells? If yes, are they reactive towards pig cells? If you do not have that information, can you tell me the epitope or peptide sequence that was used in generating that antibody? If you do not want to tell me that info, can you tell me the identity percentage of match of the pig compared to the epitope that it was used in generating the antibody. Many thanks! S

    Read More
    Answer

    Thank you for your inquiry.

    For ab758, the immunogen that was used was cytoskeletal extract of human hepatocellular carcinoma cell line, Hep3B. We have not validated this antibody in pig. But the human sequence is 85% homologous with the pig cytokeratin 8 sequence. We generally say anything with >85% homology would be expected to cross-react, so this is right on the edge. However, we do have two anti-cytokeratin 8 antibodies that we know do work in pig as ab2531 and ab2530.

    https://www.abcam.com/Cytokeratin-8-antibody-C-51-ab2531.html

    https://www.abcam.com/Cytokeratin-8-antibody-C-43-ab2530.html

    For ab27988, this antibody has been validated in pig and we therefore guarantee it will work for pig in IHC-P, IHC-Fr in accordance with our Abpromise.

    For ab7504, the immunogen was human breast carcinoma cell line, MCF-7. Pig and human EpCAM is 83% homologous, so this is a little low. We have an anti-EpCAM that is predicted (>90% homology) to cross-react with pig, and that is ab71916. However, this hasn't been validated and therefore not guaranteed in pig.

    https://www.abcam.com/EpCAM-antibody-ab71916.html

    I have to check with the lab on ab32362 and ab124687 since I don't have the immunogens readily available to BLAST.

    For ab28028, full length native protein (purified): Vimentin isolated from bovine eye lens was used as an immunogen. Pig vimentin and bovine vimentin are 97% homologous, so there is a good chance this will cross-react with pig. However this hasn't been tested and therefore isn't validated in pig. We do have a few vimentin antibodies that work in pig though,like ab20346 and ab8069.

    https://www.abcam.com/Vimentin-antibody-VI-10-ab20346.html

    https://www.abcam.com/Vimentin-antibody-V9-ab8069.html

    For ab11333, cultured human thymic fibroblasts was used as an immunogen. We don't have a particular protein sequence to look up for this one, so unfortunately we don't know how/if it will work with pig.

    I hope this information helps. I'll be in touch again once I hear about ab32362 and ab124687.

    Read More

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