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)
    

• 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)