We need to lyse the viruses for downstream immunoassays, and so it is important that the proteins are not damaged. I have only lysed cells and tissues before for SDS-PAGE, would a similar protocol with RIPA lysis be appropriate?
Protocol could be virus dependent. Treatment with appropriate detergents followed by precipitation with trichloroacetic acid. Salt treatment could also do. Please read the attached document for detail. Article Isolation of a capsid protein of bluetongue virus that induc...
https://jvi.asm.org/content/jvi/78/15/8078.full.pdf
Hi,
I believe RIPA (along with enzyme cocktails!) should be fine for western blot analysis!. However, As a thumb rule, if you're target protein is in low abundance, you might have to optimize (a bit) the initial loading volume of lysate based on the protein concentration!
I used this: https://www.scbt.com/p/ripa-lysis-buffer-system
RIPA (Radioimmunoprecipitation) Lysis Buffer System is used to lyse cells and tissue, for radio immunoprecipitation assay (RIPA).
RIPA lysis buffer has stronger denaturing capabilities than NP-40 (sc-281108) or Triton X-100 (sc-29112) and is particularly useful for disruption of nuclear membranes in the preparation of nuclear extracts.
This high quality product includes protease inhibitors, making it ready for use in mammalian cell and tissue lysis.
sc-24948, 50 mL - Components supplied in four vials:
VIAL 1: 50 mL 1X lysis buffer (pH 7.4 ±0.1)
VIAL 2: 500 μL (200mM) PMSF in DMSO
VIAL 3: 500 μL protease inhibitor cocktail in DMSO
VIAL 4: 500 μL (100mM) sodium orthovanadate in water
sc-24948A, 500 mL - Components supplied in four vials:
VIAL 1: 500 mL 1X lysis buffer
VIAL 2: 5 ml (200mM) PMSF in DMSO
VIAL 3: 5 ml protease inhibitor cocktail in DMSO
VIAL 4: 5 ml (100mM) sodium orthovanadate in water
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