FITC Anti-CD4 antibody [RPA-T4] (ab86886)

The anti-CD4 antibody [RPA-T4] (FITC), ab86886, is a Mouse IgG1. The best isotype control for ab86886 would be a Mouse IgG1 FITC conjugated such as ab18435.

ab37356 would be a good isotype control for a Mouse polyclonal IgG FITC conjugated.

Thank you very much for your inquiry.

Yes, you are absolutely correct. I can confirm, that the non- conjugated equivalent of the ab86886 is the ab86885.

When using an unconjugated antibody I can however also recommend to use an unconjugated isotype control. We forexample the ab91353as an mouse IgG1 unconjugatedisotype control.

I hopethis information is helpful. Please do not hesitate to contact us again should you haveany further question.

hank you for taking time to complete our questionnaire. I am sorry to hear that this antibody is not providing satisfactory results. The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. Having reviewed the protocol used I have a few things that I would like to clarify. The Anti-CD4 antibody ab86886 has only been tested using Flow Cytometry previously and may therefore not be suitable for use in western blotting. Have you used the antibody for analysis of cultures using Flow Cytometry to see the activity of the antibody? Additionally, you have performed the Western blot using non-reducing non-denaturing conditions. This may cause the protein to aggregate and multiple bands, make determining if the protein is detected problematic. I would suggest trying two things. 1. you could try using the antibody to detect the protein under reducing, denaturing conditions. It may be that the antibody only recognizes the native form of the protein but if it is able to detect the unfolded protein detection by Western blotting may be made easier to identify the size of the band etc. 2. you could try performing an immunoprecipitation using the antibody on non-denatures, non-reduced sample. More details of a suitable protocol can be found here: https://www.abcam.com/ps/pdf/protocols/Immunoprecipitation%20protocol%20(IP).pdf the isolated protein could then be loaded as usual on a reducing, denaturing gel for analysis. I would suggest using a positive and negative control for this if possible. You are currently using 20 ug of protein per well. It may be worth increasing this to 40-80ug if no protein is observed at the lower amount. Additionally I would suggest blocking the membrane for 1-2 hours at room temperature with 5% milk. It may also be worth trying 3% BSA as an alternative. To increase the signal it may also improve by removing the blocking agent from the diluent buffers of the antibodies, or reducing it to 0.5%. I hope these suggestions allow you to determine if the antibody is suitable for Western blotting. If you have further problems please let me know. It may be worth using an antibody such as ab51312 or ab1089 which have been testes in Western blotting previously. Or we have allophycocuanin conjugated mouse monoclonal ab18280 which may be more appropriate. I hope this information is helpful, and I thank you for your cooperation.

Thank you for contacting us. I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly. I am attaching our questionnaire so that we can gather further information regarding the samples tested and the protocol used. You do not mention wheter you have been performing Western blotting or IHC so I have attached both questionaires. Please just fill out the questionaire that is relevant to you. This should only take 5-10 minutes. Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results. This information will also allow us to investigate this case internally and initiate additional testing where necessary. If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody. I look forward to receiving your reply.