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Anti-P Glycoprotein antibody [JSB-1] (ab3366)

Reviews (1)Q&A (52)References (43)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-P Glycoprotein antibody [JSB-1] (ab3366)

    Key features and details

    • Mouse monoclonal [JSB-1] to P Glycoprotein
    • Suitable for: IHC-P
    • Reacts with: Human
    • Isotype: IgG1

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

    Product image
    Anti-P Glycoprotein antibody [EPR10364-57] (ab170904)
    • 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-P Glycoprotein antibody [JSB-1]
      See all P Glycoprotein primary antibodies

    • Description

      Mouse monoclonal [JSB-1] to P Glycoprotein

    • Host species

      Mouse

    • Specificity

      This antibody is specific for the MDR1 isoform of P Glycoprotein which is only found in the plasma membrane. It does not cross-react with MDR3. It has been shown to cross-react with Pyruvate Carboxylase, an abundant mitochondrial enzyme (130 kDa), on both immunoblots and immunohistochemical tissue sections. Weak homogeneous, cytoplasmic, or granular patterns of reactivity may represent staining of the PC cross-reactive epitope rather than positive staining for P glycoprotein.

    • Tested applications

      Suitable for: IHC-Pmore details

    • Species reactivity

      Reacts with: Human

    • Immunogen

      Multidrug-resistant Chinese hamster ovary cell line (CHrC5).

    • Epitope

      Cytoplasmic domain present on the MDR1 isoform of P Glycoprotein. This epitope is strongly conserved in mammals.

    • Positive control

      • Drug-sensitive parental cell lines and their multidrug-resistant derivatives. Human liver (positive staining detected along luminal surfaces of bilecanaliculi) or human colon (positive staining localized to luminal surface of secretaoryepithelium).

    • General notes

      This antibody is reported to work in WB according to the literature(PubMed 22389470 and 17961285). Due to customer feedback of difficulty in WB, we are no longer able to guarantee ab3366 in WB.

      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 ab3366 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
    IHC-P (1)
    1/40. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. (EDTA, pH 8-9 for example). 20-60 minutes at room temperature.
    Notes
    IHC-P
    1/40. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. (EDTA, pH 8-9 for example). 20-60 minutes at room temperature.

    Target

    • Function

      Energy-dependent efflux pump responsible for decreased drug accumulation in multidrug-resistant cells.

    • Tissue specificity

      Expressed in liver, kidney, small intestine and brain.

    • Involvement in disease

      Genetic variations in ABCB1 are associated with susceptibility to inflammatory bowel disease type 13 (IBD13) [MIM:612244]. Inflammatory bowel disease is characterized by a chronic relapsing intestinal inflammation. It is subdivided into Crohn disease and ulcerative colitis phenotypes. Crohn disease may involve any part of the gastrointestinal tract, but most frequently the terminal ileum and colon. Bowel inflammation is transmural and discontinuous; it may contain granulomas or be associated with intestinal or perianal fistulas. In contrast, in ulcerative colitis, the inflammation is continuous and limited to rectal and colonic mucosal layers; fistulas and granulomas are not observed. Both diseases include extraintestinal inflammation of the skin, eyes, or joints. Crohn disease and ulcerative colitis are commonly classified as autoimmune diseases.

    • Sequence similarities

      Belongs to the ABC transporter superfamily. ABCB family. Multidrug resistance exporter (TC 3.A.1.201) subfamily.
      Contains 2 ABC transmembrane type-1 domains.
      Contains 2 ABC transporter domains.

    • Cellular localization

      Membrane.
    • Information by UniProt

    • Database links

      • Alternative names

        • ABC20 antibody
        • ABCB1 antibody
        • ATP binding cassette, sub family B (MDR/TAP), member 1 antibody
        • ATP-binding cassette sub-family B member 1 antibody
        • CD243 antibody
        • CLCS antibody
        • Colchicin sensitivity antibody
        • Doxorubicin resistance antibody
        • GP170 antibody
        • MDR1 antibody
        • MDR1_HUMAN antibody
        • Multidrug resistance 1 antibody
        • Multidrug resistance protein 1 antibody
        • P glycoprotein 1 antibody
        • P gp antibody
        • P-glycoprotein 1 antibody
        • PGY1 antibody
        see all

      Images

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-P Glycoprotein antibody [JSB-1] (ab3366)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-P Glycoprotein antibody [JSB-1] (ab3366)
        ab3366 (4µg/ml) staining p Glycoprotein in human liver, using an automated system (DAKO Autostainer Plus). Using this protocol there is a cell membrane staining of a subpopulation of hepatocytes.
        Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.

      References (43)

      Publishing research using ab3366? Please let us know so that we can cite the reference in this datasheet.

      ab3366 has been referenced in 43 publications.

      • Niu Y  et al. Interaction of Hepatitis B Virus X Protein with the Pregnane X Receptor Enhances the Synergistic Effects of Aflatoxin B1 and Hepatitis B Virus on Promoting Hepatocarcinogenesis. J Clin Transl Hepatol 9:466-476 (2021). PubMed: 34447675
      • Vita SM  et al. P-glycoprotein Expression Is Upregulated in a Pre-Clinical Model of Traumatic Brain Injury. Neurotrauma Rep 1:207-217 (2020). PubMed: 33274346
      • Chen L  et al. Everolimus Reverses Palbociclib Resistance in ER+ Human Breast Cancer Cells by Inhibiting Phosphatidylinositol 3-Kinase(PI3K)/Akt/Mammalian Target of Rapamycin (mTOR) Pathway. Med Sci Monit 25:77-86 (2019). PubMed: 30605443
      • Koehn LM  et al. Developmental differences in the expression of ABC transporters at rat brain barrier interfaces following chronic exposure to diallyl sulfide. Sci Rep 9:5998 (2019). PubMed: 30979952
      • Hasanovic A  et al. Targeting the multidrug transporter Patched potentiates chemotherapy efficiency on adrenocortical carcinoma in vitro and in vivo. Int J Cancer 143:199-211 (2018). PubMed: 29411361
      View all Publications for this product

      Customer reviews and Q&As

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

      31-40 of 53 Abreviews or Q&A

      Question

      WB with human tissue lysate and cell lysate
      no band seen
      50 ug protein loaded
      Ab: 1/100 - 1/1000
      block: 3% milk + 2% BSA
      2nd: works well
      lot GR46599-2

      Read More
      Answer

      Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

      I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number xxx with a new lot.

      To check the status of the order please contact our Customer Service team and reference this number.

      Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

      I wish you the best of luck with your research.

      Read More

      Question

      Thank you very much for your prompt response. Yes, we would like to have one vial of ab27671 (anti-V5) for replacement. Although the Mdr1 antibody did not work well, but I am very satisfied with your help. Have a great day!

      Read More
      Answer

      Thank you for your kinds words. I am happy to help!

      As requested, I have issued a free of charge replacement for one vial of ab27671.

      To check the status of the order please contact our Customer Service team and reference this number.

      Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

      I wish you the best of luck with your research.

      Read More

      Question

      We have talked on the phone and e-mailed back and forth about the Mdr1 antibody (please see the e-mails below). We tested the replacement Mdr1 (ab3366) antibody and it still did not work. To make sure that our samples have no problem, we kept trying different Mdr1 antibody from other company. As shown on the attached picture, we did see that Mdr1 level was enhanced for A549 cells after CDDP treatment but not for pc14 cells (the right picture) when detected with one Mdr1 antibody from Santa Cruz. However, when probed with ab3366 for the same membrane, we did not see the specific signal for Mdr1 (the left picture). We would like to request replacement for anti-V5 antibody (ab27671). Hopefully this antibody will work well and I don’t need to keep bothering you. Thank you very much for your help!

      Read More
      Answer

      Thanks for your reply. I am sorry to hear the replacement vial of ab3366 did not improve things.

      Just to make sure, you would like me to send along one vial of ab27671 as a replacement for this most recent faulty vial of ab3366? If this is your request I can certainly take care of that for you.

      Read More

      Question

      I used the same protocol for all my western work. I plate 250K - 500K cells/well in 6-well plate. The next day, washed with PBS once. Added 100 uL 1xLysis buffer (diluted from 10X Cell Lysis Buffer) with 1 mM PMSF, incubated on ice for 5 min. Scraped the cells and collected them to 1.5 mL eppendorf tubes. Sonicated briefly. Spin the tube for 10 min at 13,000 g in a cold microcentrofuge. Collected supernatant.

      To prepare sample for SDS-PAGE, I used 7.5 uL 4xSDS loading buffer, 3 uL 10xDTT, 19.5 uL Lysate. Heated the sample for 10 min at 90C. Loaded all 30 uL/lane to pre-cast Tris-Glycine (4-20%) or Tris-Acetate (3-8%) SDS-PAGE. The subsequent transfer worked. Blocking buffer was TBST with 5% nonfat milk 1 hr at RT. For primary antibody (from Abcam), I tried both 1 hr at RT and overnight at 4C, none worked.

      Read More
      Answer

      Thank you for your reply.



      I am sorry that this antibody did not perform as stated on the datasheet. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used.

      Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department.

      I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

      Read More

      Question


      I used this antibody for detecting MDR1/Pgp protein in a well validated cell line. Attached please find the article where both western blot and gene expression data have shown the expression of MDR1/Pgp in this cell line.
      I repeated Western 3 times (with different antibody concentration, up to 1:100 dilution, exposure time: up to 30 min), was not able to detect MDR1 using your antibody. All my controls (beta-actin and GAPDH) worked perfectly.
      I am not satisfied with this purchase. I request a refund.

      Read More
      Answer

      According to our records, ab3366 was proving difficult to use inWB and we were in contact in order to help resolve the issue.

      Looking at our correspondence, it appears that we are awaiting more details in order to help us better understand the difficulties experienced. If the requested information has already been sent, it appears that it did not reach our Scientific Support team and we apologize for this inconvenience. In this case we would like to ask for the information again so that we can reach a resolution.

      If the issue has already been settled, please let us know so that we can be assured that the problem has been solved to your satisfaction and update our records.

      We wish you the best of luck with your research and look forward to a reply.

      Read More

      Question


      I ordered an antibody (cat#ab3366) from Abcam.
      I used this antibody for detecting MDR1/Pgp protein in a well validated cell line (KBv200). Attached please find the article where both western blot and gene expression data have shown the expression of MDR1/Pgp in this cell line.
      I repeated Western 3 times (with different antibody concentration, up to 1:100 dilution, exposure time: up to 30 min), was not able to detect MDR1 using your antibody. All my controls (beta-actin and GAPDH) worked perfectly.
      I am not satisfied with this purchase. I request a refund.

      Read More
      Answer

      Thank you for contacting Abcam regarding ab3366.

      I am sorry that you have experienced difficulties with this antibody in WB. We certainly do stand by the quality of our products and I am happy to provide a refund.


      I am hoping it would be possible to get some additional details regarding the protocol you tested, as this information will be used to further assess the quality of this product and help determine if the issue is lot specific. If you would send this to me at your earliest convenience, I would greatly appreciate it.


      Can you please confirm your order number along with the protocol information? Thank you and I look forward to your reply so that I may assist you further.

      Read More

      Question

      Last year I purchased ab3366 for use in immunocytochemistry with the HepG2 cell line (human hepatocyte carcinoma). Whilst I observe positive staining for these cells, the flourescence is always very weak when coupled to a well known secondary antibody.

      I have been using the antibody at a 1:40 dilution and incubating overnight and still the signal is weak. I perform heat retrivalwith citrate buffer.

      Please can you share any advice/recommendationsso as to increase the strength of staining?

      Read More
      Answer

      Thank you for contacting us. There are several ways you may be able to increase your signal:

      1.) How are you fixing your cells? If you are using acetone or methanol, the heat mediated antigen retrieval will not be necessary. If you are using PFA for fixation, it may help to switch to a high pH buffer such as EDTA. We have found that EDTA antigen retrieval buffer pH 8-9 to be most effective with this antibody.

      2.) It may help to use biotin - streptavidin amplification, or increase the concentration and incubation time of the secondary antibody.

      3.) What have you been using as your blocking buffer? What solution is the antibody diluted in? In some cases, using too high of a concentration of blocking agent can inhibit antibody binding.

      I hope this helps, please let me know if you need any additional assistance.

      Read More

      Question

      Catalog number(s): 3366

      Product(s): Mouse monoclonal [JSB-1] to P Glycoprotein

      Survey results
      ----------------------------------------------------------------------------------
      Telephone section
      Q. Did you contact Abcam's Scientific Support by telephone in reference to this complaint?
      A. No
      Q. Was it clear from the phone options which number you needed to press to get your issue resolved?
      A.

      Protocol advice section
      Q. Did Abcam's Scientific Support give you protocol advice?
      A.Yes
      Q. How did you feel about being given protocol advice?
      A.I requested protocol advice
      Q. Did the product work successfully after following the advice?
      A.No
      Q. Why have you not tried the protocol advice? (Select all that apply.)
      A.

      Free of charge replacement section
      Q. Did Abcam's Scientific Support give you a free of charge replacement?
      A. No

      Q. Did the replacement product work successfully?
      A.
      Credit note / refund section
      Q. Did Abcam's Scientific Support give you a credit note or a refund?
      A.No

      Q. Would you have preferred a different outcome from receiving a credit note or refund?
      A.
      Overall satisfaction section
      Q. How satisfied are you with how the complaint was handled?
      A.Satisfied
      Q. Could we have done anything to better resolve the problem?
      A.No, we found out from where came the problem.
      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
      Q. Please share with us any final thoughts you may have about Abcam, the complaints process, or our products.
      A.

      Read More
      Answer

      Vielen Dank fuer Ihre Teilnahme an Abcam's Survey.
      Bitte sagen Sie mir doch Bescheid, ob ich noch etwas fuer Sie tun kann bezueglich ab3366. Oder melden Sie sich bitte einfach falls Sie Fragen zu einem anderen Product haben.
      Viele Erfolg bei Ihren Experimenten!

      Read More

      Question

      Entschuldigung, da ist mir beim Schreiben ein Fehler unterlaufen: Fuer den
      Antikoerper PGlycoprotein (ab36743) (den es bei Ihnen nicht mehr zu
      bestellen gibt) verwendeten wir anti rabbit und fuer den Antikoerper P
      Glycoprotein (JSB-1)ab3366 (den wir jetzt hergenommen haben verwendeten wir
      anti mouse 1:5000. Auch ohne den 1. Antikoerper nahmen wir anti mouse und
      erhielten wie beschrieben unspezifische Banden.

      Read More
      Answer

      Ja, das ist kein Problem. I hatte mir schon so etwas gedacht.
      Lassen Sie mich bitte wissen, ob das Problem mit den unspezifischen Banden sich geloest hat, nachdem andere Sekundaerantikoerper ausprobiert wurden.
      Ich wuensche Ihnen viel Erfolg!

      Read More

      Question

      aus zeitlichen Gruenden konnte ich leider Ihre Fragen noch nicht beantworten.
      Wir haben einige Experimente nach Ihren Vorschlaegen durchgefuehrt. Wir haben
      die spezifische Bande erhalten, aber leider auch sehr viele unspezifische
      Banden, die fuer unser Projekt nicht gut geeignet sind.
      Zu Ihren Fragen:
      Die Spezie der Proben/Kontrollen sind human.
      Dieser CLB1 Lysis buffer ist von einer Firma entwickelt worden, von dem wir
      keine naeheren Informationen ueber die Zusammensetzung erhalten haben.
      Die Kontrolle mit Cortex extrahierten Zellen wurden mit dem Puffer 50mM
      Tris pH7,5, 150mm NaCl; 1% NP-40, 1% deoxycholate, 2% SDS and protease
      inhibitor lysiert. Hier wuerde ich erwarten, dass wir nur eine spezifische
      Bande erhalten.

      Wir hatten vorher auch RIPA-buffer zum Lysieren verwendet nach derselben
      Zusammensetzung wie in Ihrem Schreiben und anschliessend Westernblot mit dem
      Antikoerper Anti-P Glycoprotein ab36743; Referenz von 2009
      durchgefuehrt. Leider erhielten wir dann nur unspezifische Banden.
      Wir fuegten beim Transfer dem Transferbuffer auch 0,1 %SDS hinzu und
      transferierten ueber 4 Stunden, da beim Gel nach 2h-Transfer mit Coomassie
      angefaerbt noch Banden im oberen MWK zu erkennen waren.
      Nachdem wir trotzdem so viele unspezische Banden hatten, fuehrten wir ein
      Westernblott durch ohne 1. Antikoerper, nur mit dem 2. Antikoerper (10 ml
      anti rabbit HRP 1:5000 in 5% milkblock in PBS-T, 45 min 22 C) und erhielten
      ueberraschend die unspezifischen Banden.
      Alos liegt es nicht am 1. , sondern am zweiten Antikoerper.
      Vielen Dank nochmal fuer Ihre Ratschlaege.

      Read More
      Answer

      Haben Sie vielen Dank fuer Ihre email.
      Gemaess Ihrer Beschreibung scheint also der sekundaere Antikoerper die Ursache fuer die unspezifischen Banden zu sein.
      In dem Fall wuerde ich vorschlagen, einen anderen Sekundaerantikoerper zu versuchen.
      Sie haben geschrieben, dass ein anti-rabbit HRP Antikoerper verwendet wurde. Da der 1. Antikoerper in Maus erzeugt wurde (ab3366), waere ein anti-mouse HRP Antikoerper als 2. Antikoerper zu empfehlen.
      Bitte sagen Sie mir doch Bescheid, ob dass die Loesung fuer das Problem war oder ob noch immer unspezifische Banden auftreten.
      Ich wuensche Ihnen viel Erfolg!

      Read More

      31-40 of 53 Abreviews or Q&A

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