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View additional product information for M-PER™ Mammalian Protein Extraction Reagent - FAQs (78501, 78503, 78505)
16 product FAQs found
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.
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.
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.
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.
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.
M-PER Mammalian Protein Extraction Reagent is not suitable for cell fractionation. We recommend using NE-PER Nuclear and Cytoplasmic Extraction Reagents (Cat. No. 78833, 78835).
Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.
For extraction of protein from tissue samples as well as some difficult-to-lyse mammalian cells, we recommend using T-PER Tissue Protein Extraction Reagent (Cat. No. 78510) with homogenization.
Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.
If the lysis efficiency for the specific cell line is not optimal, we suggest increasing the time of incubation and the amount of M-PER reagent used. It can also help to shake the samples more
vigorously during the incubation.
Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.
The DNA would most likely be localized in the pellet but it would depend on the robustness of the nucleus in the particular cell type.
Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.
M-PER Reagent is compatible with the Thermo Scientific Coomassie Plus (Bradford) Assay (Cat. No. 23236) and the Thermo Scientific BCA Protein Assay (Cat. No. 23225) without the need to further dilute the sample for interfering substances.
Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.
Add M-PER Reagent to the cell pellet that has been washed once in PBS. Use at least 1 mL of M-PER Reagent for each 100 mg (~100 µL) of wet cell pellet. For example, 2 × 106 HeLa cells is equal to ~20 mg of cells and requires 200 µL of M-PER Reagent. Shake mixture gently for 10 minutes. Remove cell debris by centrifugation at ~14,000 × g for 15 minutes. Transfer the supernatant to a new tube for analysis.
Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.
For direct, in-plate lysis of adherent cells, protein extraction efficiency using M-PER Reagent is similar to freeze/thaw methods. For lysis of pelleted cells, either from cell suspension or scraped adherent cells, protein extraction efficiency is typically 25% higher than that achieved with freeze -thaw (three cycles) and 20% higher than sonication (2 minutes with 50% pulse) methods.
Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.
The cell lysates prepared using M-PER Mammalian Protein Extraction Reagent are compatible with many downstream assays including immunoassays, enzyme assays and a variety of common reporter assays. Additionally, because M-PER reagent is formulated with dialyzable components that are devoid of primary amines, it is more compatible with certain downstream applications, such as luciferase, beta-galactosidase, and CAT assays, than other protein extraction methods. The reagent extracts soluble proteins in a non-denatured state, enabling direct use in immunoprecipitation and other affinity purification procedures and is also compatible with Western Blot, ELISA and RIA.
Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.
M-PER Reagent efficiently extracts soluble protein from a variety of cell types, including primary cells (heart and neuron) and cells grown in suspension or adherent culture conditions (HeLa, CHO, Jurkat, NIH3T3, FM2, MCF-7, COS-7). Performance of M-PER Reagent was evaluated for yields of both total protein and specific target proteins from the various cellular compartments.
Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.
The complete cell lysis reagent contains a mild, non-denaturing detergent in 25 mM Bicine buffer (pH 7.6), that dissolves cell membranes to extract and solubilize total protein from most cellular compartments. Extraction is accomplished in only 5 minutes and requires little or no additional mechanical disruption. M-PER Reagent is formulated for minimal interference with downstream biological applications.
Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.
Please see the various lysis buffers we offer (https://www.thermofisher.com/us/en/home/life-science/protein-biology/protein-purification-isolation/cell-lysis-fractionation/cell-lysis-total-protein-extraction.html) for total protein extraction from mammalian cells.
Find additional tips, troubleshooting help, and resources within our Cell Lysis and Fractionation Support Center.