Overview

  • Product name
    Anti-cIAP2 antibody [E40]
    See all cIAP2 primary antibodies
  • Description
    Rabbit monoclonal [E40] to cIAP2
  • Host species
    Rabbit
  • Specificity
    This antibody is specific for human cIAP2. It does not cross-react with any other IAP family members.
  • Tested applications
    Suitable for: Flow Cyt, WB, IHC-P, IPmore details
    Unsuitable for: ICC
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human cIAP2 aa 400-500. The exact sequence is proprietary.

  • Positive control
    • WB: Daudi cell lysate IHC-P: Human spleen tissue, Human breast carcinoma IP: Daudi cell lysate FC: Raji cells
  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

     

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab32059 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 1/70.
WB 1/50000. Predicted molecular weight: 72 kDa.

For unpurified use at 1/1000.

IHC-P 1/600.

See IHC antigen retrieval protocols.

IP Use a concentration of 30 mg/ml.

For unpurified use at 5 µg/mL.

  • Application notes
    Is unsuitable for ICC.
  • Target

    • Function
      Apoptotic suppressor. The BIR motifs region interacts with TNF receptor associated factors 1 and 2 (TRAF1 and TRAF2) to form an heteromeric complex, which is then recruited to the tumor necrosis factor receptor 2 (TNFR2).
    • Tissue specificity
      Highly expressed in fetal lung, and kidney. In the adult, expression is mainly seen in lymphoid tissues, including spleen, thymus and peripheral blood lymphocytes.
    • Involvement in disease
      Note=A chromosomal aberration involving BIRC3 is recurrent in low-grade mucosa-associated lymphoid tissue (MALT lymphoma). Translocation t(11;18)(q21;q21) with MALT1. This translocation is found in approximately 50% of cytogenetically abnormal low-grade MALT lymphoma.
    • Sequence similarities
      Belongs to the IAP family.
      Contains 3 BIR repeats.
      Contains 1 CARD domain.
      Contains 1 RING-type zinc finger.
    • Cellular localization
      Cytoplasm.
    • Information by UniProt
    • Database links
    • Alternative names
      • AIP 1 antibody
      • AIP1 antibody
      • API 2 antibody
      • API2 antibody
      • Apoptosis inhibitor 2 antibody
      • Baculoviral IAP repeat containing 3 antibody
      • Baculoviral IAP repeat containing protein 3 antibody
      • Baculoviral IAP repeat-containing protein 3 antibody
      • BIRC 3 antibody
      • BIRC3 antibody
      • BIRC3_HUMAN antibody
      • C IAP2 antibody
      • C-IAP2 antibody
      • CIAP 2 antibody
      • CIAP2 antibody
      • HAIP 1 antibody
      • HAIP1 antibody
      • HIAP 1 antibody
      • HIAP-1 antibody
      • HIAP1 antibody
      • IAP homolog C antibody
      • IAP-1 antibody
      • Inhibitor of apoptosis protein 1 antibody
      • MALT 2 antibody
      • MALT2 antibody
      • Mammalian IAP homolog C antibody
      • MIHC antibody
      • RING finger protein 49 antibody
      • RNF49 antibody
      • TNFR2 TRAF signaling complex protein 1 antibody
      • TNFR2 TRAF signalling complex protein antibody
      • TNFR2-TRAF-signaling complex protein 1 antibody
      see all

    Images

    • Anti-cIAP2 antibody [E40] (ab32059) at 1/50000 dilution (Purified) + Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysates at 15 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 72 kDa
      Observed band size: 72 kDa

    • Anti-cIAP2 antibody [E40] (ab32059) at 1/1000 dilution (unpurified) + Daudi cell lysate.

      Predicted band size: 72 kDa
      Observed band size: 72 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast carcinoma tissue sections labeling cIAP2 with purified ab32059 at 1/600 dilution (1.12 µg/ml). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
    • Flow Cytometry analysis of Raji (Human Burkitt's lymphoma B lymphocyte) cells labeling cIAP2 with purified ab32059 at 1/70 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

    • ab32059 (purified) at 1/30 dilution (2ug) immunoprecipitating cIAP2 in Daudi whole cell lysates.
      Lane 1: Daudi (Human Burkitt's lymphoma lymphoblast) whole cell lysates 10ug
      Lane 2 (+): ab32059 & Daudi whole cell lysates
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32059 in Daudi whole cell lysates
      For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.
      Blocking and diluting buffer: 5% NFDM/TBST.

    • ab32059 (unpurified) at a 1/50 dilution staining cIAP2 in human normal spleen, using Immunohistochemistry, Paraffin Embedded Tissue.

    • cIAP2 was immunoprecipitated using 0.5mg Daudi whole cell extract, 5µg of Rabbit polyclonal to cIAP2 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
      The antibody was incubated under agitation with Protein G beads for 10min, Daudi whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
      Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab32059 (unpurified).
      Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
      Band: 72kDa; cIAP2

    References

    This product has been referenced in:
    • Lukosiute-Urboniene A  et al. Human antigen R mediated post-transcriptional regulation of inhibitors of apoptosis proteins in pancreatic cancer. World J Gastroenterol 25:205-219 (2019). Read more (PubMed: 30670910) »
    • Wang L  et al. CircDOCK1 suppresses cell apoptosis via inhibition of miR-196a-5p by targeting BIRC3 in OSCC. Oncol Rep 39:951-966 (2018). Read more (PubMed: 29286141) »
    See all 28 Publications for this product

    Customer reviews and Q&As

    1-9 of 9 Q&A

    Answer


    Please notice, these protocols may be optimised, and could not work with your specific tissue.

    Ab32059: Steam slides in 0.01M Sodium Citrate Buffer, pH 6.0 at 125°C for 30 seconds followed by 10 seconds at 90° C into a decloaking chamber / pressure cooker. Removes slides and rinse in TBST before proceeding to the staining step.

    Ab113237: Steam slides in 0.01 M sodium citrate buffer, pH 6.0 at 99-100°C - 20 minutes. Remove from heat and let stand at room temperature in buffer - 20 minutes. Rinse in 1X TBS with Tween (TBST) – 1 minute at RT.

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    Answer

    Ok great, this will ship out tonight. Please keep me updated, and I look forward to hearing from you!

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    Answer

    Thanks for getting back to me. I'm sending a vial of ab23423 free of charge on the order ***, which we have in stock and can be delivered tomorrow. Will your receiving department be open tomorrow, or should I set it for a later delivery? Please keep me updated about your coworkers' results using the older antibodies and your results with this new one. I am hopeful that one of these will work! Happy holidays and best wishes.

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    Answer

    Thanks for your reply and for your patience while i have been in touch with the lab. I have heard back from the lab, and I don't see anything unusual in their protocol so I don't have a great explanation for these results. At this point, I would suggest trying a different BIRC3 antibody (though I know you've already tried 2). I've looked through the antibodies that we have to this target, and I think that ab23423 looks promising. The WB image on the datasheet shows some extra bands (thought to be cleavage fragments or non-specific bands), but we only have 1 complaint on this antibody since it has been sold. Ab23423 has also been used in 3 references. As I mentioned before, these antibodies are covered by our guarantee, so I can issued a credit or refund if you would prefer. I apologize for all of the difficulty with these antibodies, and I'm hopeful that we can identify one that will work for you. I look forward to hearing from you. Please let me know how you would like to proceed.

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    Answer

    Thank you for your reply and for sending the additional information. I am in touch with the testing laboratory to ask for further suggestions to get the correct results. I do have a few more questions, just to make sure I understand the images that were sent. Could you possibly label the molecular weights of the ladder? And also to clarify, the left membrane with the high background is the monoclonal or the polyclonal? I apologize for the inconvenience but I have conflicting information in my notes and I want to make sure the details are accurate. I will get back to you once I hear from the lab, and please let me know if you have any questions. I look forward to hearing from you and resolving this promptly.

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    Answer

    Thanks for sending the image again. I'm sorry to see that the results are so poor with this replacement antibody. Do any of the bands on the blot correspond to the expected band size of cIAP2? How much lysate do you load into each well? The testing laboratory uses 5% non-fat dry milk instead of BSA to block the nitrocellulose membrane, and some antibodies are sensitive to blocking agent so it might be worth a try. Do you also dilute the primary and secondary antibodies in solution containing milk or BSA? This can be removed if so. Have you tried blotting for a loading control protein like beta actin on the same membrane, to check that the HRP and ECL kit are working well? We do guarantee this replacement antibody to work as well, so if the results do not improve I would be happy to issue a credit or refund for your original purchase. Please let me know if you have any questions or if there is anything else that we can do for you.

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    Answer

    Thank you for keeping me updated about this situation and for sending the protocol information and image. I unfortunately can not open the Gel file. Could you re-send it in pdf or jpg? I will be happy to see if there are any suggestions that might improve the results. The replacement antibody is also covered by our Abpromise so we would be happy to issue a credit or refund if we can not improve the results. Please let me know if you have any questions or if there is anything else you need.

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    Question
    Answer

    Thanks for your call today and for letting us know about the trouble with these antibodies. As we discussed, I am sending a free of charge replacement vial of ab113237 on the order ***, which I expect to arrive December 1. Please keep me updated about the results with ab32059 using HeLa lysate. I look forward to hearing from you. Let me know if you have any questions or if there is anything else that we can do for you. Have a great week and a happy Thanksgiving!

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    Answer

    Thank you for your phone call. I am very sorry to hear that these 3 antibodies are not providing satisfactory results. The details provided will enable us to investigate these 3 cases and will provide us with vital information for monitoring product quality. I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful. As we discussed, I believe these 3 products should have given satisfactory results. It appears that you may have received either faulty vials, or the shipment was affected by temperature variations, customs or other unknown factors. I apologize very much for the inconvenience, but am pleased to let you know that I have arranged for the replacement shipment as we discussed. Thank you for your understanding. I wish you good luck with your research! Please let me know if there is anything else I can help you with.

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