Download Cell Extraction and Lysis Extraction kits - protein

Cell Extraction and Lysis
Extraction kits - protein
NIH 3T3
M
Protein
HeLa
H
M
C6
H
M
H
Location
Flotillin
Membrane 2Spanner
COX4
Membrane 1Spanner
AchE
Peripheral
hsp90
Cytosolic
Partitioning of solubilised proteins using Thermo Scientific Mem-PER
reagent. Proteins from three cell lines were solubilised and extracted
using the Mem-PER kit. Each set of hydrophilic and hydrophobic (membrane
protein) fractions obtained were normalised to one another and analysed
by Western blot for four proteins from the cellular locations noted. The
Pierce SDS-PAGE sample prep kit was used to remove the detergent from
the membrane fraction before SDS-PAGE/Western analysis of COX4. A
negligible amount of protein was found in all debris fractions. Abbreviations:
AchE = acetylcholinesterase, COX4 = cytochrome oxidase subunit 4, hsp90
= heat shock protein 90, M = solubilised membrane protein fraction and H =
hydrophilic protein fraction.
Marker
Dpm1p
MP
PGK
VATPase
AP
M
H
M
H
M
H
Location
Integral Membrane
Protein of E.R.
Integral Membrane
Protein of Mitochondrial
Mem-PER eukaryotic membrane protein extraction kit, Thermo Scientific
Pierce
PN
No gradient method or ultracentrifugation needed.
•
Minimal cross-contamination (typically <10%) of hydrophilic protein into the hydrophobic (membrane) protein
fraction
•
Compatible with downstream applications including SDS-PAGE, Western blotting, protein assays, etc.
• Includes an easy and complete protocol for isolation of membrane proteins in approximately 1hr from mammalian
cells and tissue as well as yeast cells
• Requires only a benchtop microcentrifuge and tubes
Thermo Scientific Mem-PER eukaryotic membrane protein extraction kit, a three reagent kit enables extraction and
selective enrichment of integral and attached membrane proteins from cultured mammalian and yeast cells, as well as
from hard and soft mammalian tissues.
Catalogue No
PN89826
Cytosolic Protein
Peripheral Membrane
Protein of Yeast Vacuole
Description
Mem-PER eukaryotic membrane protein extraction kit
Includes:
Mem-PER cell lysis reagent, 10mL
Mem-PER buffer, 25mL
Mem-PER membrane protein solubilisation reagent, 40mL
Integral Membrane
of Yeast Vacuole
Use of the Thermo Scientific Mem-PER kit to solubilise and isolate
yeast integral membrane proteins (Saccharomyces cerevisiae strain
EGY-194). Approximately 10 to 15mg of wet cell paste was vortexed for 10
minutes at room temperature in 80µL of Mem-PER Reagent A and 100 to
150mg of 405 to 600 micron size acid-washed glass beads to disrupt the yeast
cell wall. A pulse spin was performed to gather the beads, and the cell lysate
was transferred to a fresh tube and kept on ice. The beads were washed with
720µL of Mem-PER Reagents B and C, and the wash was then combined with
the cell lysate and incubated on ice for 30 minutes. Samples were resolved on
4 to 12% Bis-Tris gels.
For vortex mixers and
general equipment,
please see the Fisher
Scientific Laboratory
Catalogue, or visit our
website.
See Section 4 for Cell Separation and Cellular Analysis
28
Price
180.00