Anti-P Glycoprotein antibody [EPR10363] (ab170903)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR10363] to P Glycoprotein
- Suitable for: IHC-P, WB
- Knockout validated
- Reacts with: Human
Overview
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Product name
Anti-P Glycoprotein antibody [EPR10363]
See all P Glycoprotein primary antibodies -
Description
Rabbit monoclonal [EPR10363] 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 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 & Species
See all applications and species dataApplication Species Flow Cyt HumanIHC-P HumanWB 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.
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General notes
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
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
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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
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 50% Glycerol (glycerin, glycerine), 0.05% BSA, 49% PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR10363 -
Isotype
IgG -
Research areas
Images
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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.
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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
All lanes : 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.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded Human liver tissue, labeling P Glycoprotein using unpurified ab170903 at a 1/250 dilution.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded Human kidney tissue, labeling P Glycoprotein using unpurified ab170903 at a 1/250 dilution.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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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
All lanes : Goat anti-rabbit HRP at 1/2000 dilution
Developed using the ECL technique.
Predicted band size: 141 kDa
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