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蛋白质提取和纯化（主要内容如下）
- Protein Extraction
- Protein Purification
- Protein Precipitation
- Column Preparatioin
- Q & A Posted in the Method Forum
Protein Extraction
- Whole Cell Lysate Preparation (UCSF Tumor Immunology)
For preparing proteins to be separated by IEF and SDS-PAGE, then sequenced by mass spectrometric techniques.
- Cell Membrane Preparation (John Garland)Sucrose Gradient and Centrifugation method
- Protein Extraction (Frank Bottone Jr.)
- Protein Isolation using Trizol (Frank Bottone Jr.)
- Protein Isolation From Plants (Gimila's Lab)
- Subcellular Fractionation of Proteins (PMCI)
- Protein Purification from Inclusion Bodies (Mark's Lab)
- Preparation of active proteins from inclusion bodies (NWFSC)
- Preparation of E Coli Protein for SDS-PAGE (Hahn Lab)
Simplified method for analyzing E coli protein
- Protein extracts from Aspergillus mycelium
- Phenol Extraction of Proteins (LTI)
Phenol extraction is frequently used to remove proteins from nucleic acid solutions.
Protein Purification
- Protein Purification (Michael Blaber)
Introduction to protein purification method: Initial Steps of fractionation of protein
- Protein Purification (Michael Blaber)
Guide to protein purification method: Ion exchange elutions, dialysis, concentration
? Protein Purification (Michael Blaber)
Introduction to protein purification methd: gel filtration, affinity and hydrophobic resins; preparation of resin, plumbing
? Purification of GST Fused Proteins(Contributed by Chia Jin Ngee)
Many people have vented out frustration over insoluble GST-fused proteins. This is a protocol for enzymatically active soluble GST-fused proteins. All GST-fused proteins are rendered soluble with this technique though enzyme activitiy can range from 30-90%.
? GST Fusion Protein Preparation (Vesicle Trafficking)
Detailed protocol for GST protein purification.
? Green Fluorescent Protein Isolation (Gary J. Lindquester)
? GST-Fusion Protein Purification (Mark's Lab)
? His-tag Protein Purification under Denaturing Conditions (MBS)
- PPDK His-Tag Purification (Dr. Chastain)
- PPDK De-Salting (Dr. Chastain)
- PPDK Purification via Nita Resin Column
- HeLa Cell Nuclei Preparation (John Garland)
Prepare nuclear extract from HeLa cell
? Immunopurification of Tyrosine Phosphorylated Proteins (Transduction Laboratories)
? Protein Construct Expression and Purification Procedures (Gimila's Lab)
? Chloroform Precipitation of Proteins (The Cell Biology and Cytoskeleton Group, HMS)
For concentration of proteins from a usr/localiety of aqueous solutions
? GST Fusion Protein Purification from Yeast (Dohlman Lab)
? Purification of GST Fused Proteins (PMCI)
? Sucrose Density Gradient Fractionation of Yeast Membranes (Dohlman Lab)
This is an excellent protocol for localization of membrane associated proteins in yeast.
? Purification of MBP (maLTose-Binding Proteins) Fused Proteins (PMCI)
- Purification of His-tagged Proteins from Sf9 Cells under Native Conditions (Brent Graveley)
- Making Conductivity Curves (Waters Lab)
For measuring protein conductivity during protein purification
- Purificaiton of Protein by Affinity Chromatography (Krause Lab)
Protein Precipitation
- TCA Precipitation of Proteins (Hahn Lab)
- TCA Precipitation with DOC (Waters Lab)
- Protein Precipitation (Gimila Lab)
Column Preparatioin
- Making Affinity Columns (XMMR)
- DEAE Column Preparation (Suprya Jayadev)
- DEAE column: FPLC (Suprya Jayadev)
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