What are the standard lysis buffers used with mammalian cells for detection of protein expression by immunoprecipitation (IP) or Western blot analysis?
The most commonly used buffer is RIPA Buffer with SDS. We offer RIPA Buffer (Cat. Nos. 89900 and 89901). We also offer the Pierce IP Lysis buffer (Cat. Nos. 87787 and 87788) as well as M-PER (Cat. Nos. 78501, 78503, and 78505).
Find additional tips, troubleshooting help, and resources within our Cell Lysis and Fractionation Support Center.
Is the M-PER Mammalian Protein Extraction Reagent (Cat. No. 78501, 78503, 78505) compatible with Mass spectrometry (MS)?
As any other lysis reagent, M-PER has detergent and salts in its composition, and both type of components need to be removed before the MS analysis, as they will interfere with the analysis. According to the workflow used in the MS analysis, those might be removed before the MS analysis. We recommend removing detergents at the protein level. Both detergent and salts can be removed by dialysis.
Find additional tips, troubleshooting help, and resources within our Mass Spectrometry Support Center.
Can you provide the shelf-life for the M-PER Mammalian Protein Extraction Reagent?
The M-PER Mammalian Protein Extraction Reagent is covered under our general 1-year warranty and is guaranteed to be fully functional for 12 months from the date of shipment, if stored as recommended (room temperature). Please see section 8.1 of our Terms & Conditions of Sale (https://www.thermofisher.com/content/dam/LifeTech/Documents/PDFs/Terms-and-Conditions-of-Sale.pdf) for more details.
Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.
Will M-PER Mammalian Protein Extraction Reagent extract membrane or cytoskeletal proteins?
M-PER Mammalian Protein Extraction Reagent can extract some membrane or cytoskeletal proteins, but the extraction efficiency is not consistent. The reagent was not intended to specifically extract these proteins. We recommend using Mem-PER Plus Membrane Protein Extraction Kit (Cat. No. 89842) for membrane protein extraction or Subcellular Protein Fractionation Kit for Cultured Cells (Cat. No. 78840) for compartmental extraction.
Find additional tips, troubleshooting help, and resources within our Cell Lysis and Fractionation Support Center.
When using the M-PER Mammalian Protein Extraction Reagent, is it necessary to remove cells from the plate? Should I use scraping or trypsinization to remove the cells from the plate?
M-PER reagent works very well with adherent and suspension cells, making it unnecessary to separate the cells from the plate. However, if cell removal is desired, scraping is the recommended procedure for removing cells from the plate. Trypsinization is not recommended as the free trypsin left behind in the lysis solution can damage lysed proteins.
Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.