Someone that work with human dendritic cells and western blot: how many cells I need to identify protein abundance in western blot analysis?
I want to quantify Caspase-1 and gasdermin D
it depends on expression level of your target protein in interested cell subsets, in different conditions of culture, stimuli, time etc., Fortunately, western blotting has its limitation which makes sometimes easier to choose starting concentration of protein lysate (normally 5-10ug), which means you can easily get this concentration using 2 x 10 to the 5 cells, maybe lysed in
I agree with Tuvshintugs Baljinnyam . 200`000 cells in 100uL will be fine, if possible use lover Volume +/- 50uL to be more concentrated.
I agree with Tuvshintugs Baljinnyam , for western blotting in general your focus should be more on the protein concentration of your lysate. If you obtain more than 10ug, it should be fine.
If you don't get enough protein, then try concentrating your cells as Buchs Natasha said.
I agree with the above answers, cell count of (150.000 – 250.000) will be ok, and try to concentrate your cell lysate.
- Badges
- Science method