I have this antibody solution that contains two antibody classes, IgG1κ and IgG2bκ, Can I use this as primary antibody for flow cytometry?
If so then what isotype control should I use, if needed.
Thank you.
Hello
for the part one of your question please see the attachments , for the part two of question must be to choose the proper isotype control for your experiment, match the following characteristics of the primary antibody:
Host species , Isotype , Conjugation format
Use the same experimental concentration of isotype control as primary antibody for proper interpretation of results.you can visit Abcam offers , will be given you a complete range of isotype controls, which are available both unconjugated and conjugated to: Alexa Fluor® dyes, HRP, Biotin, Phycoerythrin, FITC, Agarose, and other formats
i hope help you , Good Luck
Hello,
I wanted to add two more considerations for properly using isotypes. 1) The isotype must be at the same concentration as the primary antibodies. This is easy. 2) the isotype has to have the same F/P ratio as the primary antibody. This can be problematic. You can view more here;
http://health.usf.edu/NR/rdonlyres/57CB9476-BB36-417D-9ACE-6C77F6F6F229/27135/Isotypeeditorial.pdf
https://lists.purdue.edu/pipermail/cytometry/1998-April/009860.html
A
I think you can. The main problemis are always cross reactions. If you want to use monoclonal antibodies, you can try applying differently labeled subclasses. Another possibility is to use primary labeled antibodies from different species (one antibody could be a monoclonal, the other a polyclonal antibody or you can use two polyclonal antibodies (for instance rabbit and sheep)
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