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    products/primary-antibodies/p-glycoprotein-antibody-epr10364-ab168337.pdf

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Signal Transduction Metabolism Plasma Membrane Channels
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-P Glycoprotein antibody [EPR10364] (ab168337)

  • Datasheet
  • SDS
Reviews (2) Submit a question References (6)

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Western blot - Anti-P Glycoprotein antibody [EPR10364] (ab168337)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-P Glycoprotein antibody [EPR10364] (ab168337)
  • Western blot - Anti-P Glycoprotein antibody [EPR10364] (ab168337)
  • Western blot - Anti-P Glycoprotein antibody [EPR10364] (ab168337)
  • Western blot - Anti-P Glycoprotein antibody [EPR10364] (ab168337)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-P Glycoprotein antibody [EPR10364] (ab168337)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-P Glycoprotein antibody [EPR10364] (ab168337)
  • Anti-P Glycoprotein antibody [EPR10364] (ab168337)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR10364] to P Glycoprotein
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

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Overview

  • Product name

    Anti-P Glycoprotein antibody [EPR10364]
    See all P Glycoprotein primary antibodies
  • Description

    Rabbit monoclonal [EPR10364] 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 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.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol (glycerin, glycerine), 0.05% BSA, 59% PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR10364
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Metabolism
    • Plasma Membrane
    • Channels
    • Stem Cells
    • Hematopoietic Progenitors
    • Surface Molecules
    • Cancer
    • Drug resistance
    • P glycoproteins
    • Neuroscience
    • Processes

Associated products

  • Alternative Versions

    • Anti-P Glycoprotein antibody [EPR10364] - BSA and Azide free (ab271921)
  • Compatible Secondaries

    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • HeLa whole cell lysate (ab150035)
    • HeLa whole cell lysate (ab29545)
    • HeLa membrane extract lysate (ab29547)
    • Mouse brain tissue lysate - total protein (ab30151)

Applications

The Abpromise guarantee

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)
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 IHC antigen retrieval protocols.

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 IHC antigen retrieval protocols.

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.
  • Target information above from: UniProt accession P08183 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 5243 Human
    • Entrez Gene: 18669 Mouse
    • Entrez Gene: 24646 Rat
    • Omim: 171050 Human
    • SwissProt: P08183 Human
    • SwissProt: P06795 Mouse
    • SwissProt: P43245 Rat
    • Unigene: 489033 Human
    • Unigene: 146649 Mouse
    • Unigene: 389059 Mouse
    • Unigene: 144554 Rat
    • Unigene: 154810 Rat
    see all
  • 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

  • Western blot - Anti-P Glycoprotein antibody [EPR10364] (ab168337)
    Western blot - Anti-P Glycoprotein antibody [EPR10364] (ab168337)

    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) - Anti-P Glycoprotein antibody [EPR10364] (ab168337)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-P Glycoprotein antibody [EPR10364] (ab168337)

    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.

  • Western blot - Anti-P Glycoprotein antibody [EPR10364] (ab168337)
    Western blot - Anti-P Glycoprotein antibody [EPR10364] (ab168337)
    All lanes : Anti-P Glycoprotein antibody [EPR10364] (ab168337) at 1/2000 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 : 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.

  • Western blot - Anti-P Glycoprotein antibody [EPR10364] (ab168337)
    Western blot - Anti-P Glycoprotein antibody [EPR10364] (ab168337)
    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
    All lanes : 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.

  • Western blot - Anti-P Glycoprotein antibody [EPR10364] (ab168337)
    Western blot - Anti-P Glycoprotein antibody [EPR10364] (ab168337)
    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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-P Glycoprotein antibody [EPR10364] (ab168337)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-P Glycoprotein antibody [EPR10364] (ab168337)

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

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-P Glycoprotein antibody [EPR10364] (ab168337)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-P Glycoprotein antibody [EPR10364] (ab168337)

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

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Anti-P Glycoprotein antibody [EPR10364] (ab168337)
    Anti-P Glycoprotein antibody [EPR10364] (ab168337)

Protocols

  • Western blot protocols
  • Immunohistochemistry protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (6)

Publishing research using ab168337? Please let us know so that we can cite the reference in this datasheet.

ab168337 has been referenced in 6 publications.

  • Kinoh H  et al. Nanomedicines blocking adaptive signals in cancer cells overcome tumor TKI resistance. J Control Release 321:132-144 (2020). PubMed: 32032656
  • Sheta R  et al. Development of a 3D functional assay and identification of biomarkers, predictive for response of high-grade serous ovarian cancer (HGSOC) patients to poly-ADP ribose polymerase inhibitors (PARPis): targeted therapy. J Transl Med 18:439 (2020). PubMed: 33213473
  • Lu YL  et al. Co-delivery of Cyclopamine and Doxorubicin Mediated by Bovine Serum Albumin Nanoparticles Reverses Doxorubicin Resistance in Breast Cancer by Down-regulating P-glycoprotein Expression. J Cancer 10:2357-2368 (2019). PubMed: 31258739
  • Jiang Y  et al. The Optimization Design Of Lactoferrin Loaded HupA Nanoemulsion For Targeted Drug Transport Via Intranasal Route. Int J Nanomedicine 14:9217-9234 (2019). PubMed: 31819426
  • Ma M  et al. Celecoxib enhances sensitivity to chemotherapy drugs of T-cell lymphoma. Oncol Lett 15:4649-4656 (2018). PubMed: 29541237
  • Kang CC  et al. Single-cell Western blotting after whole-cell imaging to assess cancer chemotherapeutic response. Anal Chem 86:10429-36 (2014). Human . PubMed: 25226230

Customer reviews and Q&As

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1-2 of 2 Abreviews or Q&A

Western blot abreview for Anti-P Glycoprotein antibody [EPR10364]

Poor
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Dr. Chris Guerriero

Verified customer

Submitted Jun 27 2014

IHC - Wholemount abreview for Anti-P Glycoprotein antibody [EPR10364]

Poor
Abreviews
Abreviews
abreview image
Application
IHC - Wholemount
Sample
Zebrafish Embryo (tail region, vasculature)
Specification
tail region, vasculature
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Ryan Macdonald

Verified customer

Submitted Feb 24 2014

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