Validated using a knockout cell line
Recombinant
RabMAb

Anti-P Glycoprotein antibody [EPR10364] (ab168337)

Overview

  • Product name
    Anti-P Glycoprotein antibody [EPR10364]
    See all P Glycoprotein primary antibodies
  • Description
    Rabbit monoclonal [EPR10364] 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 ab168337 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: WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment corresponding to Human P Glycoprotein.

  • Positive control
    • HeLa, HepG2, 293T, C6 and Human fetal brain and Mouse brain lysates, Human kidney & liver tissue
  • General notes

    The mouse and rat recommendation is based on the WB results. This antibody may not be suitable for IHC with mouse or rat samples.

    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

Applications

Our Abpromise guarantee covers the use of ab168337 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 1/2000. Predicted molecular weight: 141 kDa.

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

IHC-P 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

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 - ab168337 observed at 160 kDa. Red - loading control, ab8245, observed at 37 kDa.


    ab168337 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. ab168337 and ab8245 (loading control to GAPDH) were diluted 1/500 and 1/10 000 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue sections labeling P Glycoprotein with purified ab168337 at 1/100 dilution (14 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, PH9. Hematoxylin was used to counter stain. ab97051, a Goat Anti-Rabbit IgG H&L (HRP) secondary antibody was used at 1/500 dilution. PBS instead of the primary antibody was used as the negative control.

  • All lanes : Anti-P Glycoprotein antibody [EPR10364] (ab168337) at 1/2000 dilution (purified)

    Lane 1 : Mouse brain tissue lysate
    Lane 2 : C6 (rat glioma) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG 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 and diluting buffer: 5% NFDM/TBST.

  • All lanes : Anti-P Glycoprotein antibody [EPR10364] (ab168337) at 1/2000 dilution (purified)

    Lane 1 : HeLa (human cervix adenocarcinoma) whole cell lysate
    Lane 2 : HepG2 (human hepatocellular carcinoma) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG 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 and diluting buffer: 5% NFDM/TBST.

  • All lanes : Anti-P Glycoprotein antibody [EPR10364] (ab168337) at 1/1000 dilution (unpurified)

    Lane 1 : HeLa cell lysates
    Lane 2 : 293T cell lysates
    Lane 3 : Human fetal brain lysates

    Lysates/proteins at 10 µg per lane.


    Predicted band size : 141 kDa
  • Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling P Glycoprotein with unpurified ab168337 at 1/50 dilution.

  • Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling P Glycoprotein with unpurified ab168337 at 1/50 dilution.

References

This product has been referenced in:
  • Kang CC  et al. Single-cell Western blotting after whole-cell imaging to assess cancer chemotherapeutic response. Anal Chem 86:10429-36 (2014). Human . Read more (PubMed: 25226230) »

See 1 Publication for this product

Customer reviews and Q&As

Application
Western blot
Loading amount
40 µg
Gel Running Conditions
Reduced Denaturing
Sample
Human Cell lysate - whole cell (4 different cancer cell lines muscle and blood ori)
Specification
4 different cancer cell lines muscle and blood ori
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Dr. Chris Guerriero

Verified customer

Submitted Jun 27 2014

Abcam has not validated the combination of species/application used in this Abreview.
Application
IHC - Wholemount
Sample
Zebrafish Embryo (tail region, vasculature)
Specification
tail region, vasculature
Username

Dr. Ryan Macdonald

Verified customer

Submitted Feb 24 2014

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

Sign up