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)