Hello Everyone,
I know IL-1B is a soluble protein. So how to extract it from the supernatant? Please advice. Thanks!
What is your sample? You can use RIPA lysis buffer for extraction, and then proceed with western blot protocol
https://www.hindawi.com/journals/mi/2019/7479107/
There are IL-1B antibodies available, you can use that for your western blot.
https://www.abcam.com/il-1-beta-antibody-ab2105.html
You can also use commercially available IL-1B ELISA kit
https://www.perkinelmer.com/product/alphalisa-il1-beta-kit-500-assay-pts-al220c
https://www.bosterbio.com/human-il-1-beta-picokine-trade-elisa-kit-ek0392-boster.html
Just for reference https://www.jimmunol.org/content/jimmunol/early/2017/09/13/jimmunol.1700245.full.pdf?with-ds=yes
Hello Bhairavi, my sample is in media supernatant. do i need to concentrate before adding ripa?
no need to ADD lysis buffer to conditioned media. Concentration may help. you should also be able to see it in the lysate prior to cleavage and release by Caspase 1. Remember some cells do not express caspase 1 and the IL-1 is retained in the cells.
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