Overview

  • Product name
    Anti-P Glycoprotein antibody [EPR10364-57]
    See all P Glycoprotein primary antibodies
  • Description
    Rabbit monoclonal [EPR10364-57] to P Glycoprotein
  • Host species
    Rabbit
  • 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 ab170904 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. The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat
  • Tested applications
    Suitable for: IHC-FoFr, IHC-P, WBmore details
    Unsuitable for: ICC/IF
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human P Glycoprotein aa 350-750. The exact sequence is proprietary.
    Database link: P08183

  • Positive control
    • WB: HEK-293T, C6, HeLa, HepG2 and HEK-293T cell lysates; human fetal brain tissue lysate; mouse and rat brain and kidney tissue lysates. Wild-type HAP1 cell lysate. IHC-P: Human hepatocellular carcinoma, brain and kidney tissues.
  • General notes

    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

    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

Associated products

Applications

Our Abpromise guarantee covers the use of ab170904 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-FoFr Use at an assay dependent concentration.
IHC-P 1/1200. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified use at 1/20 - 1/100

The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat

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 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

    • Lane 1: Wild type HAP1 whole cell lysate (20 µg)
      Lane 2: P Glycoprotein knockout HAP1 whole cell lysate (20 µg)
      Lane 3: HepG2 whole cell lysate (20 µg)

      Lanes 1 - 3: Merged signal (red and green). Green - ab170904 observed at 160 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab170904 was shown to specifically react with P Glycoprotein when P Glycoprotein knockout samples were used. Wild-type and P Glycoprotein knockout samples were subjected to SDS-PAGE. ab170904 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml and 1/10000 dilution respectively.

      Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    • All lanes : Anti-P Glycoprotein antibody [EPR10364-57] (ab170904) at 0.1 µg/ml (purified)

      Lanes 1 & 3 : C6 (Rat glial tumor glial cell) whole cell lysates prepared using RIPA lysis method
      Lanes 2 & 4 : C6 (Rat glial tumor glial cell) whole cell lysates prepared using 1%SDS Hot lysis method

      Lysates/proteins at 15 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) 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 and diluting buffer: 5% NFDM/TBST.

    • Immunohistochemical staining of paraffin embedded human kidney with purified ab170904 at a working dilution of 1/100. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. Counterstained 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 (inset).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue sections labeling P Glycoprotein with purified ab170904 at 1:1200 dilution (0.24 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0) ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Hematoxylinwas used as a counterstain.

      Negative control: PBS instead of the primary antibody (inset).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human hepatocellular carcinoma tissue labeling P Glycoprotein with unpurified ab170904 at 1/250 dilution.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis human kidney tissue labeling P Glycoprotein with unpurified ab170904 at 1/250 dilution.

    • All lanes : Anti-P Glycoprotein antibody [EPR10364-57] (ab170904) at 1/2000 dilution (Purified)

      Lanes 1 & 3 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates prepared using RIPA lysis method
      Lanes 2 & 4 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate prepared using 1% SDS hot lysis method

      Lysates/proteins at 15 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) 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 and diluting buffer: 5% NFDM/TBST

      Exposure time: Left image -10 seconds; Right image - 1 minute.

      We suggest not to boil the sample after lysis.

      For 1% SDS Hot Lysates preparation protocol, please refer to the protocol book in the protocol section and/or here (downloadable copy).

    • All lanes : Anti-P Glycoprotein antibody [EPR10364-57] (ab170904) at 1/2000 dilution (Purified)

      Lanes 1 & 3 : Raw264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates prepared using RIPA lysis method
      Lanes 2 & 4 : Raw264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate prepared using 1%SDS lysis method

      Lysates/proteins at 15 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) 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 and diluting buffer: 5% NFDM/TBST

      Exposure time: Left image - 3 seconds; Right image - 10 seconds.

      We suggest not to boil the sample after lysis.

    • All lanes : Anti-P Glycoprotein antibody [EPR10364-57] (ab170904) at 1/1000 dilution (unpurified)

      Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate
      Lane 2 : HepG2 (Human liver hepatocellular carcinoma cell line) cell lysate
      Lane 3 : HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) cell lysate
      Lane 4 : Human fetal brain tissue lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      Lanes 1-3 : HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution
      Lane 4 : Standard HRP labeled goat anti-rabbit at 1/2000 dilution

      Developed using the ECL technique.

      Predicted band size: 141 kDa

    • All lanes : Anti-P Glycoprotein antibody [EPR10364-57] (ab170904) at 1/1000 dilution (purified)

      Lane 1 : Mouse brain lysate
      Lane 2 : C6 (Rat glial tumor cell line) cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution

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



      Blocking/Dilution buffer: 5% NFDM/TBST.

    • All lanes : Anti-P Glycoprotein antibody [EPR10364-57] (ab170904) at 1/2000 dilution (purified)

      Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate
      Lane 2 : HepG2 (Human liver hepatocellular carcinoma cell line) cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution

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



      Blocking/Dilution buffer: 5% NFDM/TBST.

    • All lanes : Anti-P Glycoprotein antibody [EPR10364-57] (ab170904) at 1/200 dilution (unpurified)

      Lane 1 : HepG2 (Human liver hepatocellular carcinoma cell line) cell lysate
      Lane 2 : Mouse brain tissue lysate
      Lane 3 : Rat brain tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

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



      Blocking/Dilution buffer and concentration: 5% NFDM/TBST.

    • All lanes : Anti-P Glycoprotein antibody [EPR10364-57] (ab170904) at 1/1000 dilution (purified)

      Lane 1 : HepG2 (Human liver hepatocellular carcinoma cell line) cell lysate
      Lane 2 : Mouse brain tissue lysate
      Lane 3 : Rat brain tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

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



      Blocking/Dilution buffer and concentration: 5% NFDM/TBST.

    • All lanes : Anti-P Glycoprotein antibody [EPR10364-57] (ab170904) at 1/1000 dilution (unpurified)

      Lane 1 : Rat brain tissue lysate
      Lane 2 : Rat heart tissue lysate
      Lane 3 : Rat kidney tissue lysate
      Lane 4 : Rat spleen tissue lysate

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


      Exposure time: 2 minutes
    • All lanes : Anti-P Glycoprotein antibody [EPR10364-57] (ab170904) at 1/1000 dilution (unpurified)

      Lane 1 : Mouse brain tissue lysate
      Lane 2 : Mouse heart tissue lysate
      Lane 3 : Mouse kidney tissue lysate
      Lane 4 : Mouse spleen tissue lysate

      Secondary
      All lanes : HRP-conjugated goat anti-rabbit IgG (H+L) at 1/2000 dilution

      Developed using the ECL technique.

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


      Exposure time: 2 minutes
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human brain tissue labeling P Glycoprotein with unpurified ab170904 at 1/20. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody.

      Counterstained with hematoxylin.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human brain tissue labeling P Glycoprotein with purified ab170904 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody.

      Counterstained with hematoxylin.

    References

    This product has been referenced in:
    • Zhao Y  et al. The dual-inhibitory effect of miR-338-5p on the multidrug resistance and cell growth of hepatocellular carcinoma. Signal Transduct Target Ther 3:3 (2018). Read more (PubMed: 29527329) »
    • Auzmendi J  et al. Pilocarpine-Induced Status Epilepticus Is Associated with P-Glycoprotein Induction in Cardiomyocytes, Electrocardiographic Changes, and Sudden Death. Pharmaceuticals (Basel) 11:N/A (2018). IHC-FoFr ; Mouse . Read more (PubMed: 29462915) »
    See all 39 Publications for this product

    Customer reviews and Q&As

    1-7 of 7 Abreviews or Q&A

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (MDA-MB-231 cell lysate, not boiled)
    Gel Running Conditions
    Reduced Denaturing (4-20% TG gels)
    Loading amount
    40 µg
    Specification
    MDA-MB-231 cell lysate, not boiled
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

    Abcam user community

    Verified customer

    Submitted Apr 10 2018

    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (B16 tumor lysate)
    Gel Running Conditions
    Reduced Denaturing (4-20% Tris Glycin gel)
    Loading amount
    50 µg
    Specification
    B16 tumor lysate
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

    Abcam user community

    Verified customer

    Submitted Apr 10 2018

    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (Brain)
    Permeabilization
    No
    Specification
    Brain
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 12% · Temperature: RT°C
    Fixative
    Methanol followed by Acetone

    Abcam user community

    Verified customer

    Submitted Mar 15 2017

    Application
    Immunohistochemistry (PFA perfusion fixed frozen sections)
    Sample
    Rat Tissue sections (brain)
    Antigen retrieval step
    Other
    Permeabilization
    Yes - triton 1%
    Specification
    brain
    Blocking step
    Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5 · Temperature: 22°C
    Fixative
    Paraformaldehyde

    Mrs. Francoise Geffroy

    Verified customer

    Submitted Jul 06 2016

    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Breast Cancer Cells Adr)
    Gel Running Conditions
    Reduced Denaturing (10%)
    Loading amount
    20 µg
    Specification
    Breast Cancer Cells Adr
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 27°C

    Abcam user community

    Verified customer

    Submitted Jun 05 2015

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step
    peroxidase as blocking agent for 10 minute(s) · Concentration: 3% · Temperature: 25°C
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate Buffer
    Sample
    Dog Tissue sections (Kidney)
    Specification
    Kidney
    Permeabilization
    No
    Fixative
    Formalin

    Abcam user community

    Verified customer

    Submitted Sep 08 2014

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (PFA perfusion fixed frozen sections)
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Antigen retrieval step
    None
    Sample
    Rat Tissue sections (brain)
    Specification
    brain
    Permeabilization
    Yes - 0.3% Triton X-100
    Fixative
    Paraformaldehyde

    Kelly Demars

    Verified customer

    Submitted Jun 24 2013

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

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