Recombinant Anti-BCRP/ABCG2 antibody [EPR20080] (ab207732)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR20080] to BCRP/ABCG2
- Suitable for: Flow Cyt (Intra), WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-BCRP/ABCG2 antibody [EPR20080]
See all BCRP/ABCG2 primary antibodies -
Description
Rabbit monoclonal [EPR20080] to BCRP/ABCG2 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-Pmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Wild-type A549, HepG2, THP-1 cell lysate; HepG2 whole cell lysate prepared in 1% SDS Hot Lysate buffer and boiled before loading; A549 whole cell lysate prepared in RIPA lysate buffer and not boiled before loading; Human placenta lysate. IHC-P: Human placenta, liver and glioma tissues; Mouse and rat liver tissues. Flow Cyt (intra): A549 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.
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: 7.2
Preservative: 0.01% Sodium azide
Constituents: 0.05% BSA, 40% Glycerol (glycerin, glycerine), PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR20080 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab207732 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 |
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Flow Cyt (Intra) |
1/100.
ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB | (2) |
1/1000. Detects a band of approximately 75 kDa (predicted molecular weight: 72 kDa).
Sample preparation may require optimisation depending on the tissue/cell lysates. We don’t recommend this antibody for mouse and rat in WB. In our hands mouse and rat tissues showed additional bands. |
IHC-P | (1) |
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Notes |
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Flow Cyt (Intra)
1/100. ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/1000. Detects a band of approximately 75 kDa (predicted molecular weight: 72 kDa). Sample preparation may require optimisation depending on the tissue/cell lysates. We don’t recommend this antibody for mouse and rat in WB. In our hands mouse and rat tissues showed additional bands. |
IHC-P
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Target
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Function
Xenobiotic transporter that may play an important role in the exclusion of xenobiotics from the brain. May be involved in brain-to-blood efflux. Appears to play a major role in the multidrug resistance phenotype of several cancer cell lines. When overexpressed, the transfected cells become resistant to mitoxantrone, daunorubicin and doxorubicin, display diminished intracellular accumulation of daunorubicin, and manifest an ATP-dependent increase in the efflux of rhodamine 123. -
Tissue specificity
Highly expressed in placenta. Low expression in small intestine, liver and colon. -
Sequence similarities
Belongs to the ABC transporter superfamily. ABCG family. Eye pigment precursor importer (TC 3.A.1.204) subfamily.
Contains 1 ABC transmembrane type-2 domain.
Contains 1 ABC transporter domain. -
Post-translational
modificationsGlycosylation-deficient ABCG2 is normally expressed and functional. -
Cellular localization
Cell membrane. - Information by UniProt
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Database links
- Entrez Gene: 9429 Human
- Entrez Gene: 26357 Mouse
- Entrez Gene: 312382 Rat
- Omim: 603756 Human
- SwissProt: Q9UNQ0 Human
- SwissProt: Q7TMS5 Mouse
- SwissProt: Q80W57 Rat
- Unigene: 480218 Human
see all -
Alternative names
- ABC transporter antibody
- ABC15 antibody
- ABCG 2 antibody
see all
Images
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All lanes : Anti-BCRP/ABCG2 antibody [EPR20080] (ab207732) at 1/1000 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : ABCG2 knockout A549 cell lysate
Lane 3 : HepG2 cell lysate
Lane 4 : THP-1 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 72 kDa
Observed band size: 70 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab207732 observed at 70 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
ab207732 was shown to react with BCRP/ABCG2 in wild-type A549 cells in Western blot with loss of signal observed in ABCG2 knockout cell line ab259773 (ABCG2 knockout cell lysate ab259778). Wild-type A549 and ABCG2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab207732 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 h at room temperature before imaging.
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All lanes : Anti-BCRP/ABCG2 antibody [EPR20080] (ab207732) at 1/1000 dilution
Lane 1 : HepG2 (human liver hepatocellular carcinoma cell line), whole cell lysate, the lysate was prepared in 1% SDS Hot Lysate buffer and boiled before loading
Lane 2 : Huh7 (human liver hepatocellular carcinoma epithelial cell line), whole cell lysate, the lysate was prepared in 1% SDS Hot Lysate buffer and boiled before loading
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 72 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?
Exposure time: 1 minuteBlocking/Dilution buffer: 5% NFDM/TBST.
For Lysate preparation protocol, please refer to the protocol book in the protocol section and/or here (downloadable copy).
Negative control: Huh7 (PMID: 26327240, PMID: 18820285).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BCRP/ABCG2 antibody [EPR20080] (ab207732)
Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling BCRP/ABCG2 with ab207732 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on human placenta (PMID: 12237881). Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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All lanes : Anti-BCRP/ABCG2 antibody [EPR20080] (ab207732) at 1/1000 dilution
Lane 1 : A549 (human lung carcinoma cell line) whole cell lysate, the lysate was prepared in RIPA lysate buffer and boiled before loading
Lane 2 : A549 (human lung carcinoma cell line) whole cell lysate, the lysate was prepared in RIPA lysate buffer and was not boiled before loading
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 72 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?
Exposure time: 3 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
For Lysate preparation protocol, please refer to the protocol book in the protocol section and/or here (downloadable copy).
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All lanes : Anti-BCRP/ABCG2 antibody [EPR20080] (ab207732) at 1/1000 dilution
Lane 1 : Human placenta, the lysate was prepared in RIPA lysate buffer and boiled before loading
Lane 2 : Human placenta, the lysate was prepared in RIPA lysate buffer and was not boiled before loading
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 72 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?
Exposure time: 15 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
For Lysate preparation protocol, please refer to the protocol book in the protocol section and/or here (downloadable copy).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BCRP/ABCG2 antibody [EPR20080] (ab207732)
Immunohistochemical analysis of paraffin-embedded human liver tissue labeling BCRP/ABCG2 with ab207732 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on biliary canaliculis of human liver (PMID: 12237881). Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BCRP/ABCG2 antibody [EPR20080] (ab207732)
Immunohistochemical analysis of paraffin-embedded human glioma tissue labeling BCRP/ABCG2 with ab207732 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on capillaries of human glioma (PMID: 20216549). Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BCRP/ABCG2 antibody [EPR20080] (ab207732)
Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling BCRP/ABCG2 with ab207732 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on biliary canaliculis of mouse liver (PMID: 12237881). Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BCRP/ABCG2 antibody [EPR20080] (ab207732)
Immunohistochemical analysis of paraffin-embedded rat liver tissue labeling BCRP/ABCG2 with ab207732 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on biliary canaliculis of rat liver (PMID: 12237881). Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed A549 (human lung carcinoma cell line) cell line labeling BCRP/ABCG2 with ab207732 at 1/100 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (18)
ab207732 has been referenced in 18 publications.
- Tang DH et al. Inosine induces acute hyperuricaemia in rhesus monkey (Macaca mulatta) as a potential disease animal model. Pharm Biol 59:175-182 (2021). PubMed: 33715593
- Pan X et al. METTL3 promotes adriamycin resistance in MCF-7 breast cancer cells by accelerating pri-microRNA-221-3p maturation in a m6A-dependent manner. Exp Mol Med N/A:N/A (2021). PubMed: 33420414
- Sui Y et al. Roles of NANOGP8 in cancer metastasis and cancer stem cell invasion during development of castration-resistant prostate cancer. Ann Transl Med 9:45 (2021). PubMed: 33553338
- Wang C et al. Functional Doxorubicin-Loaded Omega-3 Unsaturated Fatty Acids Nanoparticles in Reversing Hepatocellular Carcinoma Multidrug Resistance. Med Sci Monit 27:e927727 (2021). PubMed: 33524008
- Sui Y et al. Insights into homeobox B9: a propeller for metastasis in dormant prostate cancer progenitor cells. Br J Cancer 125:1003-1015 (2021). PubMed: 34247196