Antibodies, Proteins, Kits and Reagents for Life Science

378 595 ₸ Product size

100 µl 378 595 ₸ 10 µl 154 118 ₸

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Anti-P Glycoprotein antibody [EPR10364-57] (ab170904)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR10364-57] to P Glycoprotein
Suitable for: IHC-P, WB
Knockout validated
Reacts with: Mouse, Rat, Human

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 & Species

Application	Species
IHC-P	Human
WB	Mouse Rat Human

See all applications and species data

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
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.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

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, 0.05% BSA, PBS
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR10364-57
Isotype

IgG
Research areas

Western blot - Anti-P Glycoprotein antibody [EPR10364-57] (ab170904)

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.
Western blot - Anti-P Glycoprotein antibody [EPR10364-57] (ab170904)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-P Glycoprotein antibody [EPR10364-57] (ab170904)

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) - Anti-P Glycoprotein antibody [EPR10364-57] (ab170904)

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) - Anti-P Glycoprotein antibody [EPR10364-57] (ab170904)

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

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-P Glycoprotein antibody [EPR10364-57] (ab170904)

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

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-P Glycoprotein antibody [EPR10364-57] (ab170904)

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).
Western blot - Anti-P Glycoprotein antibody [EPR10364-57] (ab170904)

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.
Western blot - Anti-P Glycoprotein antibody [EPR10364-57] (ab170904)

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
Western blot - Anti-P Glycoprotein antibody [EPR10364-57] (ab170904)

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.
Western blot - Anti-P Glycoprotein antibody [EPR10364-57] (ab170904)

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.
Western blot - Anti-P Glycoprotein antibody [EPR10364-57] (ab170904)

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.
Western blot - Anti-P Glycoprotein antibody [EPR10364-57] (ab170904)

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.
Western blot - Anti-P Glycoprotein antibody [EPR10364-57] (ab170904)

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
Western blot - Anti-P Glycoprotein antibody [EPR10364-57] (ab170904)

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) - Anti-P Glycoprotein antibody [EPR10364-57] (ab170904)

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) - Anti-P Glycoprotein antibody [EPR10364-57] (ab170904)

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.
Anti-P Glycoprotein antibody [EPR10364-57] (ab170904)

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