Anti-BCRP/ABCG2 antibody [BXP-21] (ab3380)

Reviews (11)Q&A (13)References (78)

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

    BXP-21 reacts with an internal epitope of BCRP/ABCG2, a 70 kD transmembrane half-transporter which is involved in Multidrug resistance. This antibody does not react with the human MDR1, MRP1 and MRP2 gene products.

  • Tested applications

    Suitable for: ICC, IHC-P, IHC-Fr, Flow Cytmore 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 cytospin preparations.
IHC-P 1/100.

Pretreated with 10 mM citric acid pH 6.0 for 20 min.
IHC-Fr 1/20.

Acetone fixed frozen sections.
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.

 

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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BCRP/ABCG2 antibody [BXP-21] (ab3380)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BCRP/ABCG2 antibody [BXP-21] (ab3380)This image is courtesy of an anonymous Abreview

      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

    • Immunocytochemistry/ Immunofluorescence - Anti-BCRP/ABCG2 antibody [BXP-21] (ab3380)
      Immunocytochemistry/ Immunofluorescence - Anti-BCRP/ABCG2 antibody [BXP-21] (ab3380)Image courtesy of Dr Gerald Davies by 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:

    • Li X  et al. NOS1 upregulates ABCG2 expression contributing to DDP chemoresistance in ovarian cancer cells. Oncol Lett 17:1595-1602 (2019). Read more (PubMed: 30675218) »
    • Koehn LM  et al. Developmental differences in the expression of ABC transporters at rat brain barrier interfaces following chronic exposure to diallyl sulfide. Sci Rep 9:5998 (2019). Read more (PubMed: 30979952) »
    See all 78 Publications for this product

    Publishing research using ab3380? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 24 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-BCRP/ABCG2 antibody [BXP-21]

     Average
    Abreviews
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (liver)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: cell conditioner 1
    Permeabilization
    No
    Specification
    liver
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Apr 09 2019

    Immunohistochemistry (Frozen sections) abreview for Anti-BCRP/ABCG2 antibody [BXP-21]

     Below Average
    Abreviews
    abreview image
    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
    Read More

    Dr. Noelia Andollo

    Verified customer

    Submitted Jun 23 2015

    Immunocytochemistry/ Immunofluorescence abreview for Anti-BCRP/ABCG2 antibody [BXP-21] - Hematopoietic/Neural Stem Cell Marker

     Good
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Dec 12 2013

    Question

    Attached is my protocol. I deparaffinized 4 times in sub-x for 5min each(total of 20min). I'm really curious to know why the dapi never stained - new bottle opened on 6/21/12. thanks for your help.

    Read More
    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

    Read More

    Question

    I wanted to make sure those above primary antibodies will work with secondary antibody Ab6785. I stained using these antibodies & nothing worked, using the strongest dilution, paraffin sections, HIER(citrate buffer Ab643214). Nothing stained not even dapi. Could the citrate buffer completely wipe out all the cells? Primary antibodies: ab3380. ab27988, ab2462, ab9221 used in conjuction with secondary antibody: ab6785. Also stained using ab52816, ab109555 with secondary Antibody ab6717. nothing stained as well, using the strongest concentration for antibodies as directed. Primary Antibody - incubation 4C overnight Secondary Antibody - incubation 1hr, RT. I want to verfiy that the secondaries to go with these primaries. I have 6 primary antibodies with 2 secondary antibodies. All single staining.

    Read More
    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

    Read More

    Question

    Our customer raise an inquiry about ab3380: Anti-BCRP/ABCG2 antibody. She would like to test in human/ EMSA. Would you please help her create the discount code?

    Read More
    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

    Question

    Some question regarding the positive control for abcam antibodies. I would like to enquire about positive control for IHC for the 3 monoclonal antibody as following: Cat No Product Name Suggested Positive control QUESTION M7802 Mo a Hu Cd44 -Can I use normal human spleen/thymus/tonsil? Ab3380 Anti-BCRP/ABCG2 antibody Caco Cell lysate Any other alternative? I don’t have this. Ab105920 Anti-ALDH1A1 THP1 and K562 cell lysate, human cerebellum and lung cancer tissue Can I use normal human liver? Hope to receive your soonest reply. Thanks Best regards,

    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

    Question

    Dear Sir/ Madam, Our customer makes a request about ab3380 (Lot NO.:GR56451-1) and ab109374(Lot NO.:GR49853-2). She has ordered these two antibodies and would like to receive the CoC. Would you please help this customer offer the CoC? Thank you very much :) Best regards,

    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

    Immunocytochemistry/ Immunofluorescence abreview for Anti-BCRP/ABCG2 antibody [BXP-21] - Hematopoietic/Neural Stem Cell Marker

     Excellent
    Abreviews
    abreview image
    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
    Read More

    Dr. Gerald Davies

    Verified customer

    Submitted Jun 03 2011

    Western blot abreview for Anti-BCRP/ABCG2 antibody [BXP-21] - Hematopoietic/Neural Stem Cell Marker

     Excellent
    Abreviews
    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
    Read More

    Ms. Rhiannon Hardwick

    Verified customer

    Submitted Jul 30 2010

    1-10 of 24 Abreviews or Q&A

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com