Overview

  • Product name
    Anti-MRP5 antibody [M5II-54]
    See all MRP5 primary antibodies
  • Description
    Rat monoclonal [M5II-54] to MRP5
  • Host species
    Rat
  • Specificity
    ab24107 does not cross-react with the human MDR1, MRP1, MRP2 or MRP3 gene products
  • Tested applications
    Suitable for: IHC-Fr, Flow Cyt, ICC/IF, IP, IHC-P, WBmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Fusion protein, corresponding to amino acids 722-910 of Human MRP5.

  • Epitope
    ab24107 reacts with an internal epitope of MRP5. ab24107 does not cross-react with the human MDR1, MRP1, MRP2 or MRP3 gene products
  • General notes


    ab24107 has potential value for detection of MRP5-mediated drug-resistance in human tumor samples

Properties

  • 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.
  • Storage buffer
    Preservative: 0.1% Sodium azide
    Constituent: 0.7% BSA

    Serum free tissue culture supernatant
  • Purity
    Tissue culture supernatant
  • Purification notes
    Antibody containing supernatant has been concentrated and filtered through a 0.22 micron filter and is serum free.
  • Primary antibody notes
    ab24107 has potential value for detection of MRP5-mediated drug-resistance in human tumor samples
  • Clonality
    Monoclonal
  • Clone number
    M5II-54
  • Myeloma
    Sp2/0
  • Isotype
    IgG2a
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab24107 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
IHC-Fr 1/20.
Flow Cyt Use 1µg for 106 cells.

ab18450 - Rat monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

 

ICC/IF 1/20 - 1/50.
IP Use at an assay dependent concentration. PubMed: 19323627
IHC-P Use at an assay dependent concentration. PubMed: 19323627
WB 1/20 - 1/50. Predicted molecular weight: 161 kDa.

Target

  • Function
    Acts as a multispecific organic anion pump which can transport nucleotide analogs.
  • Sequence similarities
    Belongs to the ABC transporter superfamily. ABCC family. Conjugate transporter (TC 3.A.1.208) subfamily.
    Contains 2 ABC transmembrane type-1 domains.
    Contains 2 ABC transporter domains.
  • Cellular localization
    Membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • ABC 33 antibody
    • ABC33 antibody
    • ABCC 5 antibody
    • Abcc5 antibody
    • ATP binding cassette sub family C (CFTR/MRP) member 5 antibody
    • ATP binding cassette sub family C member 5 antibody
    • ATP-binding cassette sub-family C member 5 antibody
    • Canalicular multispecific organic anion transporter C antibody
    • DKFZp686C1782 antibody
    • EST277145 antibody
    • MOAT C antibody
    • MOAT-C antibody
    • MOATC antibody
    • MRP 5 antibody
    • MRP5_HUMAN antibody
    • Multi specific organic anion tranporter C antibody
    • Multi-specific organic anion transporter C antibody
    • Multidrug resistance associated protein 5 antibody
    • Multidrug resistance-associated protein 5 antibody
    • pABC 11 antibody
    • pABC11 antibody
    • SMRP antibody
    see all

Images

  • Overlay histogram showing Y79 cells stained with ab24107 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab24107, 1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rat IgG (H&L) (ab98386) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rat IgG2a [aRTK2758] (ab18450, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in Y79 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

References

This product has been referenced in:
  • Zhang H  et al. Loss of miR-516a-3p mediates upregulation of ABCC5 in prostate cancer and drives its progression. Onco Targets Ther 11:3853-3867 (2018). Read more (PubMed: 30013366) »
  • Kataoka J  et al. Loss of Runt-related transcription factor 3 induces resistance to 5-fluorouracil and cisplatin in hepatocellular carcinoma. Oncol Rep 35:2576-82 (2016). IHC . Read more (PubMed: 26985715) »
See all 4 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Answer

Thank you for contacting us. As you have mentioned DAPI stains the nuclei so it helps in Immunocytochemistry to see whether the target staining observed in the images is definitely because of cells not the background staining (binding of antibody to debris). The DAPI staining should overlap with the secondary antibody staining in correct ICC/IF images. Click on the link for more ICC/IF info: https://www.abcam.com/index.html?pageconfig=resource&rid=11417 I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Answer

Thank you for contacting us. The practice of antigen retrieval is widely employed with paraffin-embedded tissue sections only.It is less often applied to cells; even in our own laboratory we generally test antibodiesby Immunohistochemistry on tissue section and by Immunocytochemistry/ Immunofluorescence on cells. The procedure we followcan be found by clicking the below given link; https://www.abcam.com/ps/pdf/protocols/icc.pdf The antigen retrieval method can also be performed with cells embedded in medium e.g paraffin. We unfortunately do not have robust protocol that we can suggest. Theoretically, any IHC protocol can be usedwith embedded cells; the protocol may however needs optimization especially the antigen retrieval step. I can provide further help on this if you could provide some details about your experience of using IHC-P and ICC/IF. The antibody ab24107 is not a purified antibody so it may contain non immune antibodies also however we do not know the concentration of these so performing ablocking step with blocking agent is recommended. Our Mitoscience colleagues have developed a protocol which you can also consider trying.Please be advised that this protocol is never tested with ab24107 so weare unable to comment how well the antibody will perform with this protocol. http://www.mitosciences.com/PDF/cyto.pdf I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Answer

Thank you for your reply and the additional information. I would like to offer a couple troubleshooting tips that might help with improving the results: 1) Using more primary antibody (maybe a 1/20 dilution) might help. 2) Trying another anigen retrieval method could improve the staining as well (e.g. Tris/EDTA pH 9 or enzymatic retrieval step). 3) Also, if human tissue sections are available to you, I would suggest trying those. The antibody was tested on human samples and is guaranteed to work on such. Should no staining be seen with human tissue, I would be able to replace or refund the antibody -since the Abpromise guarantee would apply. Should the suggestions not help with the mouse samples, I'd be happy to offer you retroactively a testing discount code, based on our testing discount program that applies for UNTESTED species and/or applications. Here is a brief description of how it works: The testing discount program is for customers who like to use an antibody/protein on an untested species/application. You would purchase the antibody at full price, test it and submit an Abreview with your data (positive or negative). On your next order you will receive a discount for ONE antibody at the full price (100%) of the antibody you have tested. The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount. I look forward to hear back from you.

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Answer

Thank you for contacting us. I am sorry to hear that this antibody is not working well. The antibody has been tested only on human samples, but not on mouse and rat. Mouse and rat are only predicted species for this antibody and we do not have any data to show if and how well it would work in these species. Therefore, we do not guarantee the antibody for predicted species. The concentration range you tried seems fine to me, although maybe a 1/20 dilution could be tried. I'd be happy to help you troubleshoot and like to know a bit more about your experiment: 1) What is the exact problem (no staining, high background)? 2) What antigen retrieval did you perform? 3) What blocking reagent was used? 4) What is being employed as detection system? Please let me know and I'd be glad to help you further. Also, IHC-P had been added to the datasheet based on this publication with PubMed: 19323627 (Karla PK et al. Expression of multidrug resistance associated protein 5 (MRP5) on cornea and its role in drug efflux. J Ocul Pharmacol Ther 25:121-32 (2009)). It might be worthwhile checking this publication for additional  protocol information as well as a control sample - although the authors used in human samples, not in mouse. I look forward to hear back from you. 

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Question
Answer

Thank you for your enquiry. The optimal dilution for FACS has not been determined for this antibody. As a general recommendation, we would suggest using 1-10ug/ml per 10^6 cells. If you have any additional questions, please do not hesitate to contact us.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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