Radioimmunoprecipitation assay buffer
Radioimmunoprecipitation assay buffer is a lysis buffer used to lyse cells and tissue for the radio immunoprecipitation assay. This buffer is more denaturing than NP-40 or Triton X-100 because it contains the ionic detergents SDS and sodium deoxycholate as active constituents and is particularly useful for disruption of nuclear membranes in the preparation of nuclear extracts. The RIPA buffer gives low background but can denature kinases.Recipe
RIPA buffer recipes vary slightly between authors and may include:
- 10-50 mM Tris-HCl, pH 7–8
- 150 mM NaCl to keep the osmotic pressure near physiological
- nonionic detergents to prevent non-specific interactions between proteins or with the tube
- anionic detergents. This needs to be optimised for every assay, the higher the concentration, the cleaner the result, but the lower the signal.
The following ingredients are optional and included as needed:
