Why reduction in protein level showed in RIPA buffer containing glycerol but not in RIPA buffer without glycerol?

10 April 2023 0 5K Report

I established a knocked down cells using shRNA and subsequently western blot to see the knock down efficiency, but two different lysate showed different result...

1st lysate

RIPA buffer + glycerol + CHAPS 1%

2nd lysate:

RIPA buffer

RIPA buffer composition (50mM TrisHCL pH8.0, 0.1% SDS, 1% NP40, 0.5% sodium deoxycholate, 150mM NaCl, 5mM EDTA)

my question why in the cell lysed with RIPA and the addition of glycerol and CHAPS, detection showed no reduction of protein in KD cells?

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