I'm doing western blot to detect LRRC8A protein which is extracted from colon cancer cell HCT116, unfortunately, I could not find a good result. Does anyone have an idea or advice?
Hi Shada,
Without seeing your results, I can just assume that your antibody is not a monoclonal antibody. Find a monoclonal antibody for your protein. When it is not the case, please share your result. If you can add one image from your western blot results, one could evaluate your problem better.
I hope this answer helps you.
Cigdem
Hi again,
I think, you need to consider following steps of your western blotting.
1- Blocking time could be longer, like overnight at 4C.
2- You can use longer washing steps between 1st and 2nd antibodies. Like 3 times 10 minutes on shaker.
4- Buy the way you can separate the proteins better if you use more suitable PAGE concentration.
Still, If you use monoclonal antibody, these could be my suggestions for you.
Good luck!
Cigdem
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