Anti-MRP5 抗体 [M5II-54] (ab24107)

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.

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.

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.

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. 

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.