Anti-Insulin Receptor beta 抗体 [18-44] (ab983)

Reviews (2)Q&A (6)References (7)
Flow Cytometry - Anti-Insulin Receptor beta antibody [18-44] (ab983)

    Key features and details

    • Mouse monoclonal [18-44] to Insulin Receptor beta
    • Suitable for: Flow Cyt
    • Reacts with: Rabbit, Cow, Human
    • Isotype: IgG2b

    製品の概要

    • 製品名

      Anti-Insulin Receptor beta antibody [18-44]
      Insulin Receptor beta 一次抗体 製品一覧

    • 製品の詳細

      Mouse monoclonal [18-44] to Insulin Receptor beta

    • 由来種

      Mouse

    • 特異性

      This antibody reacts specifically with the beta subunit of the insulin receptor.

    • アプリケーション

      適用あり: Flow Cytmore details

    • 種交差性

      交差種: Rabbit, Cow, Human

    • 免疫原

      Human placental insulin receptor.

    • 特記事項

      This product was changed from ascites to tissue culture supernatant on 19/12/2018. Please note that the dilutions may need to be adjusted accordingly. If you have any questions please do not hesitate to contact our scientific support team.

      Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

      Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

      We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

      In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

      We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

      Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

      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, as well as customer reviews and Q&As.

    製品の特性

    アプリケーション

    Our Abpromise guarantee covers the use of ab983 in the following tested applications.

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

    アプリケーション Abreviews 特記事項
    Flow Cyt Use 1µg for 106 cells.

    ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

     

    ターゲット情報

    • 関連性

      Insulin receptor mediates the biological activities of insulin by regulating multiple signaling pathways through activation of a series of phosphorylation cascades. The human insulin receptor is a heterotetrameric membrane glycoprotein consisting of disulfide-linked subunits in a ß-a-a-ß configuration. The ß-subunit (95kDa) possesses a single transmembrane domain with tyrosine kinase acivity, whereas the a-subunit (135kDa) is completely extracellular. The alpha subunits each contain insulin binding sites and are entirely extracellular in localization. The beta subunits each possess an extracellular domain, a single transmembrane domain, and a cytoplasmic tyrosine kinase domain. Binding of insulin to the alpha subunits induces a conformation change in the receptor which activates the kinase domain, stimulating tyrosine autophosphorylation of the receptor and tyrosine phosphorylation of at least five different insulin receptor substrates designated IRS-1-4, and Shc.

    • 細胞内局在

      Membrane; Single pass type I membrane protein.

    • 参照データベース

      • 別名

        • CD220 antibody
        • HHF5 antibody
        • HIR B antibody
        • INSR antibody
        • Insulin receptor antibody
        • Insulin receptor subunit beta antibody
        • IR antibody
        see all

      画像

      • Flow Cytometry - Anti-Insulin Receptor beta antibody [18-44] (ab983)
        Flow Cytometry - Anti-Insulin Receptor beta antibody [18-44] (ab983)

        Overlay histogram showing HepG2 cells stained with ab983 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab983, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HepG2 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

        This image was generated using the ascites version of the product. 

      参考文献 (7)

      ab983 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

      ab983 は 7 報の論文で使用されています。

      • Renna LV  et al. Aberrant insulin receptor expression is associated with insulin resistance and skeletal muscle atrophy in myotonic dystrophies. PLoS One 14:e0214254 (2019). PubMed: 30901379
      • Guo HH  et al. Liver-target nanotechnology facilitates berberine to ameliorate cardio-metabolic diseases. Nat Commun 10:1981 (2019). PubMed: 31040273
      • Yosten GL  et al. Evidence for an interaction between proinsulin C-peptide and GPR146. J Endocrinol 218:B1-8 (2013). ICC/IF . PubMed: 23759446
      • Thiel CT  et al. Two novel mutations in the insulin binding subunit of the insulin receptor gene without insulin binding impairment in a patient with Rabson-Mendenhall syndrome. Mol Genet Metab 94:356-62 (2008). ICC/IF ; Human . PubMed: 18411068
      • Brindle NP  et al. Anti-(insulin receptor) monoclonal antibody-stimulated tyrosine phosphorylation in cells transfected with human insulin receptor cDNA. Biochem J 268:615-20 (1990). PubMed: 1694662
      • Prigent SA  et al. Identification of epitopes on the human insulin receptor reacting with rabbit polyclonal antisera and mouse monoclonal antibodies. J Biol Chem 265:9970-7 (1990). PubMed: 1693619
      • Soos MA  et al. Monoclonal antibodies reacting with multiple epitopes on the human insulin receptor. Biochem J 235:199-208 (1986). PubMed: 2427071

      レビューと Q&A

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      1-8 of 8 Abreviews or Q&A

      Western blot abreview for Anti-Insulin Receptor beta antibody [18-44]

       Good
      Abreviews
      abreview image
      Application
      Western blot
      Sample
      Rabbit Recombinant protein (NA)
      Gel Running Conditions
      Reduced Denaturing (Bio Rad 4-15% Criterion)
      Loading amount
      1 µg
      Specification
      NA
      Blocking step
      BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
      Read More

      Makenzie Woodford

      Verified customer

      投稿 Oct 11 2019

      Western blot abreview for Anti-Insulin Receptor beta antibody [18-44]

       Excellent
      Abreviews
      abreview image
      Application
      Western blot
      Loading amount
      15 µg
      Gel Running Conditions
      Non-reduced Denaturing (10)
      Sample
      Mouse Tissue lysate - other (Hep G2)
      Specification
      Hep G2
      Blocking step
      BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
      Read More

      Abcam user community

      Verified customer

      投稿 Jan 27 2014

      Question


      I would like to ask if you offer the insulin receptor antibody specific
      for exon 11 (aa 745-756). The goal should be to distinguish between IR-A
      and IR-B isoform of insulin receptor. Exon 11 is specific for IR-B
      isoform. Or if you offer alternative how to "catch" specifically IR-B
      isoform of insulin receptor.
      Thank you very much.
      Yours sincerely,

      Read More
      Answer



      According to the Uniprot homepage the alpha chain is located from amino acid 28-758, whereas the beta chain is from aa 763-1382:

      http://www.uniprot.org/uniprot/P06213

      ab69508 has its immunogen in the cytoplasmic domain of the beta subunit; this sequence seen here http://www.uniprot.org/uniprot/P06213#PRO_0000016687 as aa 980-1382 is buried in the beta subunit and does not fall in the alpha subunit which is aa 28-758 so I would not expect cross reactivity with alpha in this case. This has not been experimentally tested.

      https://www.abcam.com/index.html?datasheet=69508 (or use the following: https://www.abcam.com/index.html?datasheet=69508).

      ab983 : I am happy to confirm that product ab983 is specific for the β-subunit of human insulin receptor. This was determined by (1) immunoblotting on human insulin receptor and (2) epitope analysis of short receptor peptides. The minimum epitope for this antibody was defined as a sequence PEEHRP corresponding to amino acids 765-770 close to the N-terminus of the β-subunit.

      https://www.abcam.com/index.html?datasheet=983 (or use the following: https://www.abcam.com/index.html?datasheet=983).

      ab80527 : This antibody was generated using synthetic peptide corresponding the aa Tyr-KKNGRILTLPRSNPS from the C-terminal of human insulin receptor B-subunit. And therefore is also expected to be specific for the beta chain.

      https://www.abcam.com/index.html?datasheet=80527 (or use the following: https://www.abcam.com/index.html?datasheet=80527).

      More information on these antibodies can be found on their respective datasheets by following the links.

      Read More

      Question

      Post complaint survey:
      ...
      Q. Could we have done anything to better resolve the problem?
      A.The original CD63 antibody did not work despite the trying various permutations of blocking reagents etc. I requested an alternative antibody TSG101 (ab983)and this was offered free of charge to compensate for the original CD63. The TSG101 was used on saliva samples and has been published. I managed to get a positive immunosignal however it was comparatively weak to the published data even though I had followed the paper protocol. The data sheet that accompanied the antibody did not have an immunoblotting protocol. Please could abcam suggest a protocol as the aliqout was small to start with so I would like to avoid repeating the immunoblotting. KInd regards xx
      Q. Would you mind if our scientific support team contacted you to better understand what happened and reach an agreeable solution?
      A.No, I would not mind
      ...

      Read More
      Answer

      Thank you very much for having provided your feedback in the last survey.

      I am happy to hear that you were satisfied with our service and that the ab83 does work better than the original antibody received. I am sorry though, that the ab83 still does not work to your entire satisfaction. The laboratory used a classical protocol with 5% milk blocking to test this antibody. Would you like to troubleshoot the protocol together? I am happy to look into troubleshooting the ab83. If you would like our help, please provide the additional information on the samples and the protocol used with an image inthe attached questionnaire.

      Thank you for your cooperation. Please do not hesitate to contact us, either with the information, or also if you have any other question or concern.

      Read More

      Question

      What is the epitope of this antibody? Do you know if it binds C-terminally?

      Read More
      Answer

      Thank you for your inquiry and your patience with this reply. We have limited data regarding the specific epitope for this antibody, but it appears that it is amino acids 765-770 (PEEHRP). I hope this information helps. Please contact us with any other questions.

      Read More

      Question

      How are you all, love your team, anyways i have a customer which used your product and all failed on WB & IP. She would like to receive credit on this, can you credit our account? below is the protocol I had her email to me as i know this is required for your team to see if followed procedure correctly. Please let me know ASAP! Thanks (word per word from customer) I used Cat. No.AB983, Lot #101002 both in an immunoprecipitation experiment and a Western blot. It did not work in either application. Immunoprecipitation Protocol (from another company) 1. Add 1 mg of cell lysate at a concentration of roughly 1µg/µl total cell protein to a microcentrifuge tube. 2. Pre-clean the lysate by adding 60µl (30µl packed beads) of washed Protein A agarose bead slurry (Catalog # 16-125). Gently rock at 4°C for 1 hour. 3. Collect the agarose beads by pulsing (5 seconds in the microcentrifuge at 14,000 x g), and collect the supernatant. 4. Added 10 µl of anti-Insulin Receptor, Ab983 to the supernatant. 5. Gently rock the reaction mixture at 4°C overnight. 6. Capture the immunocomplex by adding 60µl of washed Protein A agarose bead slurry (30µl packed beads). 7. Gently rock the reaction mixture at 4°C for 1-2 hours. 8. Collect the agarose beads by pulsing (5 seconds in the microcentrifuge at 14,000 x g), and drain off the supernatant. Wash the beads 4 times with ice-cold cell lysis buffer. 9. Resuspend the agarose beads in 60µl 2X Laemmli sample buffer. 10. boil at 95°C for 5 minutes. 11. Collect the beads after boiling using a microcentrifuge pulse. 12. Perform SDS-PAGE and immunoblot analysis on a sample of the supernatant fraction. WESTERN: Used AB983 at a 1:1000 concentration in BSA Binding Buffer. No detection. [Another company's ab worked great] I would love to receive credit on the credit card for this purchase. Thanks.

      Read More
      Answer

      Thanks for your email and I'm sorry to hear that your customer is experiencing difficulty with this antibody. I do require some additional details in order to investigate this. Exactly what type of samples were used? And from which species? How was the sample prepared? For the Western, how much protein was loaded? Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.)? How long was the primary incubation period? What were the blocking conditions and for how long? What was the secondary antibody, concentration, and incubation period? Also, with the other company's antibody that was used - was it a Insulin Receptor beta antibody? Thank you and I look forward to hearing from you.

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      Question

      I want to know how good the reactivity is with bovine (cow) species as it was mentioned in the product data sheet. Could you please send me any western blot pictures or any other data showing the reactivity of this antibody in bovine species? Thank you in advance and hoping to hear from you at your earliest.

      Read More
      Answer

      Thank you for your enquiry. The reactivity of this antibody with bovine and rabbit insulin receptor is extremely weak. In Western blots, a very faint band can be seen in some samples, but this banding is not seen consistently in all samples. We mention this weak cross-reactivity in our data sheet in case this is potentially problematic for users of this antibody, but we do not recommend this antibody for detecting bovine insulin receptor.

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      Question

      What is the reactivity of the above antibody in bovine samples? On the datasheet it states "weak reactivity, but what does the WB look like and what dilution is needed. ?

      Read More
      Answer

      Thank you for your enquiry. This antibody produces very faint banding when used at relatively high concentration (25 ug/ml) to immunoblot bovine or rabbit insulin receptor. In contrast, immunoblotting on human insulin receptor produces a sharp, strong band. We do not recommend this antibody for detection of bovine insulin receptor, but we wanted to bring this weak level of cross-reactivity to customers' attention in case it could pose a problem in someone's assays.

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