Description

This product offers a convenient and efficient method for isolating both cell membrane and cytoplasmic proteins from animal cells or tissues. The extracted membrane protein fraction includes not only plasma membrane proteins but also those integral to the membranes of mitochondria, the endoplasmic reticulum, and the Golgi apparatus. This kit uses a new and highly efficient detergent formulation. First, a mild detergent is used to permeabilize cells and release soluble cytoplasmic proteins. Then, another detergent is used to dissolve membrane proteins. The extraction efficiency depends on the number of times the integral membrane protein(s) span the lipid bilayer. For proteins with at least one or two transmembrane domains, the extraction efficiency is usually as high as 90%. The cross-contamination of cytoplasmic proteins into the membrane fraction is usually less than 10%. Selective detergent extraction eliminates the need for phase separation based on hydrophobicity, allowing for better reproducibility and higher throughput.

Features

Unparalleled Yield:

Extracting approximately 0.4 to 0.6 mg of membrane protein from 5×10^6 cells, this formulation consistently outperforms competing products by 30% to 50%.
Exceptional Purity:

With a cross-contamination rate of less than 10%, this method ensures minimal cytosolic protein contamination, resulting in exceptional purity of the extracted membrane protein fraction.
Simplified Sample Preparation::

Cell samples do not require homogenization, freeze-thaw cycles, or other complex pretreatment procedures.
High Compatibility:

Membrane proteins can be extracted from animal cells and tissues and subsequently used in experiments such as BCA protein quantification, SDS-PAGE, Western blot, and immunoprecipitation.
Fast Speed:

Membrane protein extraction can be completed within about 90 minutes.

Outperforms competing products with a higher yield of target protein

Fig 1. Four different types of membrane proteins (with monomer molecular weights of 70.6 kDa, 78.8 kDa, 103.6 kDa, and 39.8 kDa respectively) were overexpressed in HEK293F cells. Equal amounts of cells (5 × 10^6) were taken and membrane proteins were extracted using two different kits. WB detection was performed using Anti-Strep tag II antibody. Lysates at 10 µl per lane.

Fig 1. Four different types of membrane proteins (with monomer molecular weights of 70.6 kDa, 78.8 kDa, 103.6 kDa, and 39.8 kDa respectively) were overexpressed in HEK293F cells. Equal amounts of cells (5 × 10^6) were taken and membrane proteins were extracted using two different kits. WB detection was performed using Anti-Strep tag II antibody. Lysates at 10 µl per lane

Excellent membrane fractionation fficiency—with minimal cytosolic protein contamination in the membrane protein fraction

Fig. 2. The membrane protein (M) and cytosolic protein (C) components were separated from HEK293F cells using Thermo 89842 and YamayBio BF7475 kits. WB detection was performed using an Anti-HSP 90 antibody. Lysates at 10 µl per lane.

Fig. 2. The membrane protein (M) and cytosolic protein (C) components were separated from HEK293F cells using Thermo 89842 and YamayBio BF7475 kits. WB detection was performed using an Anti-HSP 90 antibody. Lysates at 10 µl per lane.

Highly adaptable to different cell lines with a 30-50% higher yield

Fig 3. Take 5 × 10^6 cells of different types respectively, and use the kits of Thermo 89842 and YamayBio BF7475 to extract membrane proteins. The extracted membrane proteins were quantified by BCA protein quantification kit (e.g., Cat No. BH5484).

Fig 3. Take 5 × 10^6 cells of different types respectively, and use the kits of Thermo 89842 and YamayBio BF7475 to extract membrane proteins. The extracted membrane proteins were quantified by BCA protein quantification kit (e.g., Cat No. BH5484).

Efficiently isolated membrane proteins from tissue samples

Fig 4. Membrane proteins were extracted from 20 mg of mouse heart tissue using the YamayBio BF7475 protocol. The extracted membrane and cytosolic protein samples were then subjected to Western blot analysis using various target antibodies.

Fig 4. Membrane proteins were extracted from 20 mg of mouse heart tissue using the YamayBio BF7475 protocol. The extracted membrane and cytosolic protein samples were then subjected to Western blot analysis using various target antibodies.

Specifications

Product Type	Protein Extraction Kit
Form	Liquid
Sufficient For	10 Samples / 50 Samples
Reagent Type	Cell Wash Buffer / Cell Permeabilization Buffer / Cell Solubilization Buffer
Quantity	60 mL / 300 mL

Storage

Upon receipt aliquot and store Cell Permeabilization Buffer and Solubilization Buffer at -20°C to avoid freeze/thaws. Store Cell Wash Solution at 4°C. Product is shipped on ice.

Documents

BF7475-10 COA

BF7475-50 COA

SDS

User Guide

Flyer of Product_ Animal Membrane and Cytosol Protein Extraction Kit

Animal Mammalian Membrane and Cytosol Protein Extraction Kit-Yamaybio