I am using the Mem-PER Plus Membrane Protein Extraction Kit with less than 5 X 10E6 cells. Should I scale down the amount of reagents accordingly?
The kit reagents can be scaled up or down as long as the ratio of reagents to cells is maintained. However, we don't have specific guidelines for the range of cell numbers that can be used because protein yields can vary depending on the cell line. It is important to note that the process of scaling the kit reagents up or down and determining the appropriate cell numbers for optimal protein yield will require empirical optimization by the investigator.
Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.
What kind of detergent does the Permeabilization Buffer from the Mem-PER Plus Membrane Protein Extraction Kit (Cat. No 89842) contain?
The composition of the Permeabilization Buffer, including the identity and concentration of any detergent therein, is proprietary. However, the Permeabilization Buffer contains a very small amount of a weak non-ionic detergent to facilitate the permeabilization of the cells and release of the cytosolic proteins.
Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center
Will the Mem-PER Plus Membrane Protein Extraction Kit (Cat. No. 89842) extract membrane proteins from the plasma membrane only or also from other intra-cellular membranes (e.g., nuclear, mitochondrial, endoplasmic reticulum)?
The Permeabilization buffer in the Mem-PER Plus Membrane Protein Extraction Kit (Cat. No. 89842) creates pores in the plasma membrane of cells that releases cytosolic proteins. The permeabilized cells are centrifuged, and the supernatant contains the cytosolic protein fraction. The pellet is then resuspended in the Solubilization buffer containing a proprietary detergent that solubilizes plasma, mitochondrial, and ER membrane proteins. Subsequent centrifugation yields a supernatant containing solubilized membrane and membrane-associated proteins.
Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center
Can microalgae be processed with the Mem-PER Plus Membrane Protein Extraction Kit (Cat. No. 89842)?
The Mem-PER Plus Membrane Protein Extraction Kit is not suitable for any organism that has a cell wall, including algae, plants, yeast, or bacteria.
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.