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)