Overview

  • Product name
    Anti-BCRP/ABCG2 antibody [BXP-21]
    See all BCRP/ABCG2 primary antibodies
  • Description
    Mouse monoclonal [BXP-21] to BCRP/ABCG2
  • Host species
    Mouse
  • Specificity
    This antibody does not react with the human MDR1, MRP1 and MRP2 gene products.
  • Tested applications
    Suitable for: ICC, IHC-P, IHC-Fr, Flow Cyt, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Fusion protein composed of E. Coli maltose binding protein and BCRP peptide [126 amino acids, 271-396 AA of BCRP, GeneBank accession #AF098951].

  • Positive control
    • IHC-P: Human breast cancer tissue. ICC/IF: MCF7 cells. Caco Cell lysate (see second reference cited).
  • General notes

    For the WB, please note the subcellular localization and distribution of the BCRP/ABCG2 in Normal Human Tissues: PMID:11309308

    We have received mixed feedback from customers, the antibody showed 45-50 kDa band size in A549 and Caco2 cell lines against predicted size of 72kDa. In response to data we have received we no longer guarantee this antibody for western blot. ABCG2 is a multi-pass membrane protein so for troubleshooting we would recommend heating the samples at 60-70C for 15-20 minutes instead of boiling and as this protein exists as homodimer so complete reduction of samples is highly recommended. In case of any other question please email scientific support team.

Properties

Applications

Our Abpromise guarantee covers the use of ab3380 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
ICC 1/20 - 1/50. Acetone fixed cells.
IHC-P 1/100.
IHC-Fr 1/20 - 1/40.
Flow Cyt Use at an assay dependent concentration. PubMed: 24136221

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

 

ICC/IF Use at an assay dependent concentration.

Target

  • 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
    modifications
    Glycosylation-deficient ABCG2 is normally expressed and functional.
  • Cellular localization
    Cell membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • ABC transporter antibody
    • ABC15 antibody
    • ABCG 2 antibody
    • ABCG2 antibody
    • ABCG2_HUMAN antibody
    • ABCP antibody
    • ATP binding cassette sub family G (WHITE) member 2 antibody
    • ATP binding cassette transporter G2 antibody
    • ATP-binding cassette sub-family G member 2 antibody
    • BCRP antibody
    • BCRP1 antibody
    • BMDP antibody
    • Breast cancer resistance protein antibody
    • CD338 antibody
    • CDw338 antibody
    • CDw338 antigen antibody
    • EST157481 antibody
    • GOUT1 antibody
    • MGC102821 antibody
    • Mitoxantrone resistance associated protein antibody
    • Mitoxantrone resistance-associated protein antibody
    • MRX antibody
    • Multi drug resistance efflux transport ATP binding cassette sub family G (WHITE) member 2 antibody
    • MXR antibody
    • MXR1 antibody
    • Placenta specific ATP binding cassette transporter antibody
    • Placenta specific MDR protein antibody
    • Placenta-specific ATP-binding cassette transporter antibody
    • UAQTL1 antibody
    see all

Images

  • Immunohistochemical analysis of human breast cancer tissue, staining BCRP/ABCG2 with ab3380.

    Tissue was fixed with formaldehyde, blocked with 3% H2O2 for 5 minutes at 20°C and permeabilized with PBS-Tween; antigen retrieval was by heat mediation in a citrate buffer (pH 6). Samples were incubated with primary antibody (1/200 in PBS-Tween) for 1 hour at 20°C. A biotinylated goat anti-mouse polyclonal IgG was used as the secondary antibody.

    See Abreview

  • ab3380 staining BCRP/ABCG2 in MCF7 human breast cancer cells by Immunocytochemistry/ Immunofluorescence.
    Cells were fixed in paraformaldehyde and permeabilized using 0.1% Triton X-100. Samples were then blocked using 10% serum for 2 hours at 25°C, then incubated with ab3380 at a 1/50 dilution for 12 hours at 4°C. A rabbit anti-mouse polyclonal conjugated to Alexa Fluor 594 was used as the secondary at a 1/50 dilution.

    See Abreview

References

This product has been referenced in:
  • Szabó E  et al. A new fluorescent dye accumulation assay for parallel measurements of the ABCG2, ABCB1 and ABCC1 multidrug transporter functions. PLoS One 13:e0190629 (2018). Read more (PubMed: 29342177) »
  • Naruse T  et al. Lower gingival squamous cell carcinoma with brain metastasis during long-term cetuximab treatment: A case report. Oncol Lett 15:7158-7162 (2018). Read more (PubMed: 29725438) »
See all 66 Publications for this product

Customer reviews and Q&As

1-10 of 23 Abreviews or Q&A

Application
Immunohistochemistry (Frozen sections)
Sample
Rabbit Tissue sections (Rabbit cornea. Limbal epithelial area. Only progen)
Specification
Rabbit cornea. Limbal epithelial area. Only progen
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde

Dr. Noelia Andollo

Verified customer

Submitted Jun 23 2015

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
Sample
Human Cell (ES-derived hepatocytes)
Specification
ES-derived hepatocytes
Permeabilization
Yes - BSA diluted in PBS/0.1% Tween
Fixative
Methanol

Abcam user community

Verified customer

Submitted Dec 12 2013

Answer

Thank you for your reply and these additional details.

When you write "I deparaffinized 4 times in sub-x for 5min each" I am unsure what you mean by sub-x. We recommend the following depariffinization protocol:



1. Xylene: 2 x 3 minutes

2. Xylene 1:1 with 100% ethanol: 3 minutes

3. 100% ethanol: 2 x 3 minutes

4. 95% ethanol: 3 minutes

5. 70 % ethanol: 3 minutes

6. 50 % ethanol: 3 minutes

7. Running cold tap water to rinse

Barring this issue, we can see no reason why you should be having no staining with any of these product or with a Dapi stain. Have you attempted using positive control tissues for these products?

I look forward to your reply. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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Answer

Thank you for contacting us.

I am very sorry to hear of the troubles which you have been having in IHC. You have indeed matched the corrected primary and secondary antibodies for this application. Antigen retrieval using a citrate buffer should not cause this effect. I am curious as to why Dapi,a fluorescent stain that binds strongly to A-T rich regions in DNA, would not be working for you either. Could you please send to me the details of your protocol including all sample preparation steps which you have taken? It seems possible to me that you are seeing incomplete de-paraffinizing or that you may have issues with the fixation of these tissues.

I have included in this email our IHC-P protocol and our IHC troubleshooting guide for your review as well.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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Answer

Thank you for contacting us. We do not accept reviews of EMSA at this time, so I am not able to offer the testing discount for this application.

The antibody is capable of recognizing BCRP/ABCG2 in its native, 3D conformation, as suggested by the ICC and IHC data. It will probably be useful for EMSA of native samples too, but we do not guarantee this.

I am sorry we cannot provide a discount code in this instance. I wish your customer good luck in her research.

Read More

Answer

Thank you for contacting us. ab3380; The ABCG2 target is expressed in tonsils, spleen, gall bladder and lungs etc so the sections of these tissues will be good positive control. You can find the complete expression information on Human Protein Atlas website; http://www.proteinatlas.org/ENSG00000118777/normal ab4372; ab87930 are spleen tissue slides ab4349 is human lung tissue slide ab105920; normal liver sections can be used; this target is expressed in normal liver. http://www.proteinatlas.org/ENSG00000165092/normal ab4348 is normal liver tissue slide. M7802; this does not seems to be Abcam product however CD44 is highly expressed in spleen, tonsils etc so the tissue sections of these will be a good positive control. http://www.proteinatlas.org/ENSG00000026508/normal I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information. Check the Abcam range of IHC kits also; https://www.abcam.com/index.html?pageconfig=resource&rid=12906 https://www.abcam.com/index.html?pageconfig=resource&rid=12853 https://www.abcam.com/index.html?pageconfig=resource&rid=12886

Read More

Answer

Thank you for your enquiry. Please find enclosed the two requested CoC for ab3380 (Lot NO.:GR56451-1) and ab109374 (Lot NO.:GR49853-2) as attachments. I hope this helps and if I can assist further, please do not hesitate to contact me.  

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (MCF7 human breast cancer cells)
Specification
MCF7 human breast cancer cells
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton X-100 in PBS
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C

Dr. Gerald Davies

Verified customer

Submitted Jun 03 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Tissue lysate - whole (Liver)
Loading amount
50 µg
Specification
Liver
Gel Running Conditions
Non-reduced Denaturing (7.5%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 27°C

Ms. Rhiannon Hardwick

Verified customer

Submitted Jul 30 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Cancer cell line)
Loading amount
15 µg
Specification
Cancer cell line
Gel Running Conditions
Reduced Denaturing (7.5%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 27°C

Abcam user community

Verified customer

Submitted Aug 07 2009

1-10 of 23 Abreviews or Q&A

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