Validated using a knockout cell line
Recombinant
RabMAb

Anti-P Glycoprotein antibody [EPR10363] (ab170903)

Overview

  • Product name
    Anti-P Glycoprotein antibody [EPR10363]
    See all P Glycoprotein primary antibodies
  • Description
    Rabbit monoclonal [EPR10363] to P Glycoprotein
  • Specificity
    P-glycoprotein 1 (also known as Multidrug resistance protein 1) has a predicted molecular weight of 141 kDa, however it has 3 potential glycosylation sites (N-linked) which may affect the migration of the protein. In our hands ab170903 detects a predominant protein band migrating in the region of 180-200 kDa and typically will demonstrate a smear on the membrane in the region of the 150 – 300 kDa due to the glycosylation profile of the protein. It may be necessary to optimise your cell or tissue lysis protocol to efficiently extract P-glycoprotein 1 as it is a multi-pass membrane protein. Abcam recommends not boiling the sample after lysis.
  • Tested applications
    Suitable for: IHC-P, WBmore details
    Unsuitable for: Flow Cyt or ICC/IF
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Recombinant fragment within Human P Glycoprotein aa 950 to the C-terminus. The exact sequence is proprietary.
    Database link: P08183

  • Positive control
    • HeLa, HepG2 and 293T cell lysates; Human kidney, liver and uterus tissues; 293T cells.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    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.

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

Properties

Applications

Our Abpromise guarantee covers the use of ab170903 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 1/250 - 1/600. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB 1/1000 - 1/5000. Predicted molecular weight: 141 kDa.

For optimal detection Abcam recommends not boiling the sample after lysis.

  • Application notes
    Is unsuitable for Flow Cyt or ICC/IF.
  • 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



    • Predicted band size : 141 kDa

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: P glycoprotein knockout HAP1 cell lysate (20 µg)
      Lane 3: HeLa cell lysate (20 µg)
      Lane 4: HepG2 cell lysate (20 µg)

      Lanes 1 - 4: Merged signal (red and green). Green - ab170903 observed at 160 kDa. Red - loading control, ab8245, observed at 37 kDa.


      ab170903 was shown to recognize P glycoprotein when P glycoprotein knockout samples were used, along with additional cross-reactive bands. Wild-type and P glycoprotein knockout samples were subjected to SDS-PAGE. ab170903 and ab8245 (loading control to GAPDH) were diluted 1/500 and 1/10000 respectively and incubated overnight at 4°C. 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 1/10000 dilution for 1 hour at room temperature before imaging.

    • Immunohistochemical staining of paraffin embedded human kidney with purified ab170903 at a working dilution of 1/600. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
    • All lanes : Anti-P Glycoprotein antibody [EPR10363] (ab170903) at 1/10000 dilution (purified)

      Lane 1 : HepG2 cell lysate
      Lane 2 : HEK293 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution

      Predicted band size : 141 kDa
      Observed band size : 180 kDa (why is the actual band size different from the predicted?)

      Blocking buffer: 5% NFDM/TBST
      Dilution buffer: 5% NFDM/TBST

    • Immunohistochemical analysis of paraffin-embedded Human uterus tissue, labeling P Glycoprotein using unpurified ab170903 at a 1/250 dilution.

    • Immunohistochemical analysis of paraffin-embedded Human liver tissue, labeling P Glycoprotein using unpurified ab170903 at a 1/250 dilution.

    • Immunohistochemical analysis of paraffin-embedded Human kidney tissue, labeling P Glycoprotein using unpurified ab170903 at a 1/250 dilution.

    • All lanes : Anti-P Glycoprotein antibody [EPR10363] (ab170903) at 1/1000 dilution (unpurified)

      Lane 1 : HeLa cell lysate
      Lane 2 : HepG2 cell lysate
      Lane 3 : 293T cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      Goat anti-rabbit HRP at 1/2000 dilution
      Developed using the ECL technique

      Predicted band size : 141 kDa

    References

    This product has been referenced in:
    • Xiang C  et al. Pulmonary expression of CYP2A13 and ABCB1 is regulated by FOXA2, and their genetic interaction is associated with lung cancer. FASEB J 29:1986-98 (2015). Read more (PubMed: 25667220) »

    See 1 Publication for this product

    Customer reviews and Q&As

    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    Western blot
    Sample
    Guinea pig Tissue lysate - whole (Intestine, Liver, Brain, LS180 cell lysate)
    Gel Running Conditions
    Non-reduced Denaturing (8 % gel, Samples were either boiled 5 min at 95°C or not)
    Loading amount
    30 µg
    Specification
    Intestine, Liver, Brain, LS180 cell lysate
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    Submitted Apr 19 2014

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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