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Validated using a knockout cell line

Anti-MRP2 antibody [M2III-5] (ab15603)

Submit a review Q&A (3)References (10)
Western blot - Anti-MRP2 antibody [M2III-5] (ab15603)

    Key features and details

    • Mouse monoclonal [M2III-5] to MRP2
    • Suitable for: WB
    • Knockout validated
    • Reacts with: Human
    • Isotype: IgG2b

    Overview

    • Product name

      Anti-MRP2 antibody [M2III-5]
      See all MRP2 primary antibodies

    • Description

      Mouse monoclonal [M2III-5] to MRP2

    • Host species

      Mouse

    • Specificity

      This antibody does not cross-react with the human MDR1 P-gp, MRP1, MRP3, or MRP5 gene products.

    • Tested applications

      Suitable for: WBmore details

    • Species reactivity

      Reacts with: Human
      Does not react with: Mouse

    • Immunogen

      This antibody was raised against a fusion protein of the bacterial maltose binding protein and rat MRP2, containing the 202-amino acid C terminal end of the transporter protein.

    • Epitope

      ab15603 reacts with an internal epitope of MRP2; a 190-200 kD transmembrane protein earlier known as the canalicular multi-organic anion transporter cMOAT, absent in patients with the Dubin-Johnson syndrome.

    • 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

    Applications

    The Abpromise guarantee

    Our Abpromise guarantee covers the use of ab15603 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
    WB
    1/20 - 1/50. Predicted molecular weight: 185.4 kDa.
    Notes
    WB
    1/20 - 1/50. Predicted molecular weight: 185.4 kDa.

    Target

    • Function

      Mediates hepatobiliary excretion of numerous organic anions. May function as a cellular cisplatin transporter.

    • Tissue specificity

      Found on the apical membrane of polarized cells in liver, kidney and intestine. The highest expression is found in liver.

    • Involvement in disease

      Defects in ABCC2 are the cause of Dubin-Johnson syndrome (DJS) [MIM:237500]. DJS is an autosomal recessive disorder characterized by conjugated hyperbilirubinemia, an increase in the urinary excretion of coproporphyrin isomer I, deposition of melanin-like pigment in hepatocytes, and prolonged retention of sulfobromophthalein, but otherwise normal liver function.

    • Sequence similarities

      Belongs to the ABC transporter superfamily. ABCC family. Conjugate transporter (TC 3.A.1.208) subfamily.
      Contains 2 ABC transmembrane type-1 domains.
      Contains 2 ABC transporter domains.

    • Cellular localization

      Membrane.
    • Information by UniProt

    • Database links

      • Alternative names

        • ABC30 antibody
        • abcC2 antibody
        • ATP binding cassette sub family C (CFTR/MRP) member 2 antibody
        • ATP binding cassette subfamily C member 2 antibody
        • ATP-binding cassette sub-family C member 2 antibody
        • Canalicular multidrug resistance protein antibody
        • Canalicular multispecific organic anion transporter 1 antibody
        • CMOAT antibody
        • CMOAT1 antibody
        • cMRP antibody
        • DJS antibody
        • KIAA1010 antibody
        • MRP 2 antibody
        • MRP2_HUMAN antibody
        • Multidrug resistance associated protein 2 antibody
        • Multidrug resistance-associated protein 2 antibody
        see all

      Images

      • Western blot - Anti-MRP2 antibody [M2III-5] (ab15603)
        Western blot - Anti-MRP2 antibody [M2III-5] (ab15603)
        All lanes : Anti-MRP2 antibody [M2III-5] (ab15603) at 1/20 dilution

        Lane 1 : Wild-type A549 (Human lung carcinoma cell line) whole cell lysate
        Lane 2 : ABCC2 knockout A549 (Human lung carcinoma cell line) whole cell lysate

        Lysates/proteins at 20 µg per lane.

        Performed under reducing conditions.

        Predicted band size: 185.4 kDa



        Lanes 1 - 2: Merged signal (red and green). Green - ab15603 observed at 210 kDa. Red - loading control, ab52866, observed at 50 kDa.

        ab15603 was shown to recognize ABCC2 in wild-type A549 cells as signal was lost at the expected MW in ABCC2 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and ABCC2 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab15603 and ab52866 (Rabbit anti-alpha Tubulin loading control) were incubated overnight at 4°C at 1/20 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

      References (10)

      Publishing research using ab15603? Please let us know so that we can cite the reference in this datasheet.

      ab15603 has been referenced in 10 publications.

      • Yang AJ  et al. Acute exercise and brain BACE1 protein content: a time course study. Physiol Rep 7:e14084 (2019). PubMed: 31033251
      • Baranowski BJ  et al. Evaluation of neuropathological effects of a high-fat high-sucrose diet in middle-aged male C57BL6/J mice. Physiol Rep 6:e13729 (2018). PubMed: 29890051
      • Kataoka J  et al. Loss of Runt-related transcription factor 3 induces resistance to 5-fluorouracil and cisplatin in hepatocellular carcinoma. Oncol Rep 35:2576-82 (2016). IHC . PubMed: 26985715
      • Hardwick RN  et al. Molecular mechanism of altered ezetimibe disposition in nonalcoholic steatohepatitis. Drug Metab Dispos 40:450-60 (2012). WB ; Rat . PubMed: 22112382
      • Saeki J  et al. LPS-induced dissociation of multidrug resistance-associated protein 2 (Mrp2) and radixin is associated with Mrp2 selective internalization in rats. Biochem Pharmacol 81:178-84 (2011). WB, IHC-Fr ; Rat . PubMed: 20868650
      • Hardwick RN  et al. Variations in ATP-binding cassette transporter regulation during the progression of human nonalcoholic fatty liver disease. Drug Metab Dispos 39:2395-402 (2011). WB ; Human . PubMed: 21878559
      • Sekine S  et al. Oxidative stress is a triggering factor for LPS-induced Mrp2 internalization in the cryopreserved rat and human liver slices. Biochem Biophys Res Commun 399:279-85 (2010). IHC-Fr ; Human, Rat . PubMed: 20655877
      • Ogura J  et al. Alteration of Mrp2 and P-gp expression, including expression in remote organs, after intestinal ischemia-reperfusion. Life Sci 82:1242-8 (2008). WB ; Rat . PubMed: 18538350
      • Yang XX  et al. A mechanistic study on reduced toxicity of irinotecan by coadministered thalidomide, a tumor necrosis factor-alpha inhibitor. J Pharmacol Exp Ther 319:82-104 (2006). WB ; Rat . PubMed: 16815871
      • Scheffer GL  et al. Specific detection of multidrug resistance proteins MRP1, MRP2, MRP3, MRP5, and MDR3 P-glycoprotein with a panel of monoclonal antibodies. Cancer Res 60:5269-77 (2000). PubMed: 11016657

      Customer reviews and Q&As

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

      1-3 of 3 Abreviews or Q&A

      Question

      I have a question concerning the antibody ab15603 [M2III-5] MRP-2.

      I have seen that some publications used an antibody from other companies like Enzo identified M2III-5. It was used to detect the MRP-2 protein in mouse. On the Enzo website, it is indicated that this clone react with human, rat and mouse. However, on your website, the same clone number is indicated to not react with mouse.

      I want to know if the clone numbers (ex: M2III-5) are in fact representing the exact same thing between every companies?

      I hope that my question is clear.

      Read More
      Answer

      Thank you for contacting us.

      The note regarding mouse specificity is based on data from the following reference, which includes a description of the clone's development and characterization:

      Cancer Res. 2000 Sep 15;60(18):5269-77.
      PMID: 11016657
      http://cancerres.aacrjournals.org/content/60/18/5269.full.pdf+html

      Table 1 has a summary of species reactivity for this clone and several others.

      Further characterization may have shown reactivity with other murine samples, in other applications, and this may account for the information on the datasheets of other vendors, but we do not have this data.

      It is possible that there is more than one clone raised against MRP2 designated "M2III-5" but this seems less likely than the possibility of differences in testing conditions and samples across different laboratories.

      I hope this is helpful. Please do not hesitate to contact us if you need any more information.

      Read More

      Question

      which MRP2 antibody would be the best to test against PIG?

      Read More
      Answer

      Thank you very much for your inquiry and your patience while waiting for our answer.

      I have inquired with the laboratories of the polyclonal antibodies, whether the immunogen sequence is conserved with the pig sequence. This would then provide a good basis for testing the antibody.

      The ab110740 has unfortunately only 79%homology with the pigsequence and we can not recommend it therefore.

      Also for the ab58709, the alignment is only 62%, and we can not recommend it neither. I am very sorry.

      For the monoclonal antibodies, it is more difficult to predict, as the epitopes have not been mapped. As however the overall sequence alignment is relatively good, they might have also good changes to crossreact with the pig protein. Please see here the overall alignment: http://www.ebi.ac.uk/Tools/services/web_clustalw2/toolresult.ebi?tool=clustalw2&jobId=clustalw2-I20120627-135954-0702-94637034-oy

      The ab3373 and the ab15603 have been raised against the human protein sequence 1339-1541. There are many conserved residues.
      The ab24106 and the ab130385 have been raised against the human sequence 860-950, which seems less conserved than the C terminal. The ab3372 has been raised against human sequence 215 to 310, again, a less conserved region than the C terminus.

      So in summary, I would think either the ab3373 or ab15603 most likely to crossreact with the pig protein from the choice of antibodies we have. This has however not been tested, and it can well be that they will not react with the pig protein.
      https://www.abcam.com/MRP2-antibody-M2-III-6-ab3373.html (or use the following: https://www.abcam.com/MRP2-antibody-M2-III-6-ab3373.html).
      https://www.abcam.com/MRP2-antibody-M2III-5-ab15603.html (or use the following: https://www.abcam.com/MRP2-antibody-M2III-5-ab15603.html).

      If you would therefore like to test this, I can offer a discount off a future purchase if you buy ab3373 or ab15603 now, test it inpig and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of: 1 free primary antibody.

      If you are interested in this offer, please follow these steps:

      1. Reply to this e-mail to let me know that you would like to proceed and test ab3373 or ab15603 in pig. I will then send a discount code. This code must be issued before purchasingthe antibodyso please wait for my reply before ordering.

      2. Purchase ab3373 or ab15603 either by phone, fax, or online (www.abcam.com).

      3. Test it in pig.

      4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

      5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for anyprimary antibodyordered and the discount code is valid for 4 months after issue.
      Please remember that submission of the Abreview is sufficient for the discount code to become active.

      We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab3373 or ab15603 turns out to be unsuitable for pig, you will still receive the discount on your next purchase after your Abreview has been submitted.

      Please let me know if you have any questions about this offer and I would be happy to help you further.

      The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

      Read More

      Question

      I have used your M2III-5 antibody to examine the expression of mouse Mrp2 protein. Your product info sheet stated that M2III-5 can cross-react with rat, mouse and human Mrp2. Strangely, on our western blot a 170KD band was shown in both the wild type and the Mrp2 knockout samples. Honestly, I would think that band is the Mrp2 band if I did not have the Mrp2 KO sample as a negative control. We have work with M2III-6 a lot in our research, M2III-6 work beautifully in the WT and Mrp2-/- rats. From literature, Scheffer GL et al developed the clone for M2III-5 antibody and in their paper "M2III-5 was shown NO cross-reactivity to the mouse Mrp2" (table 1 of the paper, Cancer Research 2000, 60:5269-77). Also from the paper, M2III-5 and M2III-6 recognize the same aa sequence (1336-1541). M2III-6 has no cross-reactivity with mouse Mrp2 either although both M2III-5 and 6 can react with rat Mrp2 very well. I wonder if there is any difference between your M2III-5 antibody and the authors'. From my conversation with one of your tech support person this morning, you use the same clone as the authors. I personally think they should have the same reactivity. Could you please explain me why your antibody has different reactivity from author's? Is there any possibility that your M2III-5 can recognize a band other than Mrp2 in the mouse sample? Have you confirmed the specificity of your M2III-5 in mouse samples? P.S. attachment of M2III-5 results with moue liver sample.

      Read More
      Answer

      Thank you for your feedback regarding this product. As a result of the information that you have sent to me and references sourced via “Pub Med” I have currently removed cross reactivity with from our product data base. I would like to thank you for the information that you have forwarded and would be more than willing to offer either a credit note to your account/ refund. Could you confirm for me the order details cited below and I will arrange a credit note to be issued? Your PO or our Abcam order number The name of your purchasing agent and the shipping address. I look forward to hearing from you soon.

      Read More

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