Anti-CD3 antibody [PS1] (ab699)

Thank you for contacting us. I'm sorry that the staining currently isn't working as you would like. There are a few suggestions that may help the results. Firstly I came across this description of how to prepare frozen sections. I thought it quite interesting (if a little wordy) but particularly in regards to the importance in quickly preparing samples to achieve good, specific staining. http://www.ihcworld.com/_protocols/histology/frozen_section_technique_1.htm Additionally, as you say it would be worth trying a few different dilutions for the primary antibodies. Although counter intuitively I would suggest trying higher concentrations: 1/50, 1/100, 1/250 as these antibodies are not very highly concentrated. Another thing that may improve the non-specific binding considerably is introducing a separate blocking step. Try incubating the sample with 10% serum from species which secondary antibody was raised in or 3% BSA in PBST for 30 min. I would also see if the incubation time with the antibody has an effect, try both 1 hour at room temperature and as you are doing, overnight at 4°C. I would continue to use 1% BSA in both the primary and secondary antibody diluent as you are doing currently. With washing using PBS between each step. As you have performed a "no primary" control, were the results of this clean? I was a little confused as to what secondary antibodies you are using. Would you be able to supply the catalog numbers (even if from other companies). Before carrying out the double staining I would try to optimise the staining for each antibody then combine the protocols after this has been achieved. I hope this information has been helpful. If you have any further questions please do not hesitate to ask. I look forward to hearing if there are any improvements.

Thank you for your enquiry. I can confirm that ab699 CD3 antibody [PS1] is a mouse monoclonal IgG2a antibody. We have the following isotype control, suitable for flow cytometry: ab91361 Mouse IgG2a [ICIGG2A] - Isotype Control Tested and guarnateed in Flow Cyt Click here (or use the following: https://www.abcam.com/Mouse-IgG2a-Kappa-Monoclonal-B128-Isotype-Control-ab91361.html). I can recommend to review the datasheet for further information. If you have any further questions, please do not hesitate to contact us.

Thank you for your enquiry and your interest in our products. The isotype of this antibody is mouse IgG2a. You can conduct a search based on the isotype using our recently developed search engine at this site: https://www.abcam.com/index.html?c=718 I have conducted a very quick search for you and found some products which may be suitable for you. Could you please take a look at the on-line datasheets to see which one would suit your need. -ab1281, -ab91361, -ab91364, -ab91362 -ab91365 -ab91363 I hope this helps and if I can assist further, please do not hesitate to contact me.