How to choose flow cytometry antibodies

Antibodies are an important part of many flow cytometry experiments. In this video, we take a look at the structure of antibodies, the way they interact with cells during staining, and how to select them.

[@jfromtheusa]

so helpful, amazing first stop for help on this!

[@ajarieger_flow]

Thank you! 😊

[@QNNDukeNguyen]

Yea agree!

[@debajitbhowmick7079]

Hi
Just discovered your channel, pretty cool. Never seen such good video, audio, and graphics used in any other videos. At 8:18 you mentioned the term "best". How we can define the "best" most logically possible? Just having a high frequency of positive signals can not the only parameter. Especially in the case of antigens that are not regular or intracellular targets, the access of which is very much dependent on fix/perm protocol.

[@ajarieger_flow]

Thanks for your feedback and comment! In this case, if I were to define "best", it would be based on which antibody best allowed me to discriminate my population of interest, keeping in mind the biology I expect. This is an interplay between the antibody and the fluorochrome you have chosen to pair it with.

[@debajitbhowmick7079]

@@ajarieger_flow do you have some case studies or different examples to show why one clone is better than another? wrong clones can show a huge positive population. It becomes more problematic when the biology is not well known.

[@ajarieger_flow]

@@debajitbhowmick7079 This is best done on an experiment by experiment basis.

[@fatima.Hubaishi]

Hi Aja,
Thank you I love your videos. I heard that there is a limit up to three cytokines for Abs when I do ICS? is it true?

[@ajarieger_flow]

Thanks for tuning in! I have not ever heard this before and have done more than 3 myself. The number you can do will depend on the configuration of your cytometer- but will also be affected by fluorochrome available (tandem dyes may not always work nicely for intracellular staining) and the kinetics of expression of what you want to look at (i.e. are the cytokines expressed with similar timing). Please reach out if you have any other questions :)

[@fatima.Hubaishi]

@@ajarieger_flow Thank you so much! I get your point!!