Plant organelle/membrane isolation
- Mitochondrial fraction
- Nuclear fraction
- Plasma membrane fraction
- Thylakoid extraction
- Arabidopsis thylakoid preparation
- Arabidopsis thylakoid extraction
- Arabidopsis lumen extraction
- BBY preparation
- Chlorophyll measurements
- PSII RC extraction for cryo-EM
- Extraction of leaf proteins
- Diatom protein extraction
- Phenol protein extraction
- Protein extraction from grasses
- Ponceau membrane staining
- TCA acetone precipitation method
- Western blot protocol
- Western blot video tutorial
- Peptide neutralization/competition assay
- Simultaneous Western blot
- Quantitative Western blot
- Quantitative Western blot video tutorial
- Western blot troubleshooting
- Western blot using IgY
- Dot blot
- ELISA
- Immunoprecipitation
- Immunoprecipitation/IgY
- Immunohistochemistry
- Anti-KLH antibody removal
- Yolk delipidation
Technical information
Antibody typesPurification
- Antibody purification
- Antibody purification - small amount of protein
- IgY purification methods
- Protein purification using antibodies
- Elution of antibodies from affinity columns
Protocols > Plasma membrane fractionProtocol for the isolation of plasma membranes from plant tissues. Protocol courtesy of Dr. Masayoshi Maeshima, Laboratory of Cell Dynamics, Graduate School of Bioagricultural Sciences Nagoya University, Nagoya, Japan. In the original protocol, 2-4 mg from 200 g of mung bean hypocotyls of 3.5-day-old seedlings were homogenized and used. The plasma membrane accounts for 10% of the total membrane fraction (crude membrane fraction). The ER membrane is rich in the crude membrane fraction. This protocol can be scaled-down. If 50-100 g of plant tissue is used, the first precipitate should be suspended with 2 ml of sucrose/KP/DTT, followed by mixing the suspension with 2 ml of the phase separation solution. Protocol
|
PEG 3540 (SIGMA P-3640) | 3.0 g | |
DextranT500 (Pharmacia) | 3.0 g | |
Sucrose/KP/DTT | 28.0 ml | |
300 mM NaCl/sucrose/KP/DTT | 4.8 ml |
IMPORTANT: Store at 0°C (on ice) in four 50 ml Falcon tubes.
STOCK SOLUTIONS
1. 100 mM K-phosphate buffer, pH 7.8 (200 ml)
2. 0.25 M sucrose, 10 mM K-phosphate buffer, pH 7.8, 1 mM DTT (sucrose/KP/DTT) (500 ml)
3. 300 mM NaCl in sucrose/KP/DTT (100 ml)