I recently started working with pigs, and unfortunately the library of fluorochrome-conjugated antibodies for flow is not at all as large as it is for humans or mice, so I have wondered if I could use some of the Abs marketed for other applications instead. Has anyone experience with this?
Thanks in advance
I have tried some ICC antibodies for flow cytometry and they worked very well provided the dilution factor is less in case of flow cytometry eg. if you are using a 1:200 dilution for ICC then you need to use some 1:100 dilution for flow cytometry. In my case it worked very well. but it may vary in other cases. All the best..
It is possible to do it, you are just going to have to do a titration of both the primary and secondary antibody and make sure you have all the appropriate controls when you are testing it out. Like Ajay said, you're likely going to have to use your antibody in a less dilute form, so make sure when you test it out you are doing a wide range of dilutions to make sure you're not missing out. Held og lykke med det! :)
Completely agree with the above replies. Also look how the antibody was used in ICC in particular, if there was a fixation step you will likely need a similar fix step in your flow cytometry workflow to ensure similar epitope presentation. With that said it is of course worth trying the antibodies out with live cells in case you need to perform a full FACS in future.
I hope they work out for you.
The short answer is certainly, but the long answer is maybe. If you're interested in a surface protein, you need to make sure that your antibody has specificity to an extracellular epitope. Another concern is that if your antibody isn't directly conjugated, you're going to have to do two step staining (primary antibody followed by a fluorochrome). While this this is easy enough to do, it sometimes adds a lot of background which can make FACS analyses difficult. As the others suggested, you're really gonna have to do this on a case by case basis for each antibody of interest. You'll need to titer the amount (preferably on cells that you know express the antibody, and maybe even a negative control cell) as well as use appropriate isotype and/or FMO controls. Best of luck!
Hello Thomas,
Just wanted to add further, recently CST has introduced an antibody range named XP monoclonal antibodies, in which you can use an antibody for any type of application. You can just try..!!
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