蛋白质提取和纯化

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)
    
• (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.

·         (Gary J. Lindquester)

·         (Mark's Lab)

·         His-tag Protein Purification under Denaturing Conditions (MBS)

• (Dr. Chastain)
      
• (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
    
• (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)