U-Blot™ RIPA Lysis Buffer

RIPA (Radio-Immunoprecipitation Assay) buffer has been used for protein extraction from mammalian cells and tissues. The buffer contains low concentration of ionic and non-ionic detergents, which usually do not interfere with antigen-antibody binding.  The protocol is relatively simple and extracted proteins can be used in applications such as Western blotting, ELISA and immunoprecipitation.  Although RIPA buffer has been used widely, it comes with following disadvantages:

1. The protein profile extracted with RIPA buffer is incomplete.
2. Protein loss to the pellet fraction is non-proportional, non-selective and unpredictable.
3. Many artifacts have been reported using RIPA for apoptosis studies.
4. Due to loss of proteins, the amount and ratio of proteins extracted are deviated from those    actually present in the cells or tissues resulting in biased data interpretation especially when quantitative and semi-quantitative experiments are involved.