I'm preparing for two protein immunoblot assay from cell lysate.
The reference what i referred used two lysis buffer for cell lysis.
For immunoprecipitation before western blot, NETN buffer (TBSN; 50 mM Tris, 100 mM NaCl, 1.0% Nonidet P-40, pH 7.5) or (20 mM Tris-HCl [pH 7.4], 100 mM NaCl, 1 mM EDTA, 0.5% NP-40).
But just for immunoblot, RIPA buffer (50 mM Tris–HCl, pH 8.0, 150 mM NaCl, 1% NP-40, 0.5% sodium deoxycholate, 0.1% SDS) or (50 mM Tris-HCl, 150 mM NaCl, 5 mM EDTA, 0.5% NP-40) or (50 mM Tris, 150 mM NaCl, 0.1% SDS, 0.5% deoxycholic acid, 1% NP40, pH 7.4) etc.
I want to ask what is the difference betwwen RIPA and NETN buffer usage for cell lysis.
RIPA buffer and NETN buffer are both commonly used lysis buffers for the extraction of proteins from cells, but they have different compositions and properties.
RIPA buffer (Radioimmunoprecipitation assay buffer) is a strong lysis buffer that is commonly used for the extraction of proteins from mammalian cells and particularly useful for the extraction of membrane-bound and nuclear proteins, and it can be used to extract both cytoplasmic and nuclear proteins from cells. However, due to its strong lysis properties, it may also solubilize non-specific proteins and interfere with downstream applications such as immunoprecipitation.
NETN buffer (Nuclear and cytoplasmic extraction buffer) is a mild lysis buffer that is used to separate the nuclear and cytoplasmic fractions of cells. NETN buffer is particularly useful for the extraction of nuclear proteins, as it separates the nuclear and cytoplasmic fractions of cells. However, it may not be as effective at extracting membrane-bound proteins or other cytoplasmic proteins.
In summary, RIPA buffer is a strong lysis buffer that is useful for the extraction of both membrane-bound and nuclear proteins, while NETN buffer is a mild lysis buffer that is useful for the separation of nuclear and cytoplasmic fractions of cells. The choice of buffer will depend on the specific experimental requirements and the type of proteins that need to be extracted.
- Badges
- Science topic
- Similar topics
- Gels