蛋白質電泳

蛋白質電泳（主要內容如下）

• One-Dimensional SDS-PAGE

• Two-Demensional SDS-PAGE

• Protein Electrophoresis in Agarose Gel 

• Gel Staining

• Recipes

One-Dimensional SDS-PAGE

·         Protein Gel and Staining (Gottschling Lab)
Provides procedures for gel preparation, gel staining...

·         Preparation of SDS-Polyacrylamide Gels (SDS-PAGE) (William H. Heidcamp)

·         SDS Gel Electrophoresis (Petra Klaff)
Resolving and stacking gel

·         SDS-Page Gel Electrophoresis (Dr. Chastain)

·         SDS PAGE Gels (Gimila Lab)

·         SDS-Polyacrylamide gels (NWFSC)
Standard Laemmli protocol

·         Denaturing Discontinuous Polyacrylamide Gel Electrophoresis (SDS-PAGE)(Goldberg Lab)
Very nice and detailed protocol

• PolyAcrylamide Gel Electrophoresis (PAGE) gels
  Preparation of stock solutions of acrylamide and method of preparation of convex,  and gradient gels
    

·         SDS-PAGE Gels (PMCI Research)
Detailed protocol for preparing gel, reagents for SDS-PAGE and mini-gels

·         Tricine-Polyacrylamide Gels (NWFSC)
For high resolution of small proteins

·         Tricine SDS-PAGE (Goldberg Lab)
Detailed protocol  

·         Electroblotting of SDS-PAA gels (Molecular Genetics Network Lab)

• SDS-polyacrylamide gels (single or double Minigel system) (Molecular Genetics Network Lab)
    

·         Electroelution of Proteins From SDS-PAGE Gels (Mike A. Dyer)
This is a general protocol that was developed for the ISCO electroeluter but could easily be applied to other systems.

Two-Demensional SDS-PAGE

·         2-D PAGE Protein Analysis (ExPASy)

o        Analytical 2-D PAGE protocols

o        Preparative 2-D PAGE protocols

o        Post-separation analysis  

·         Isoelectric Focusing as the First Dimension (UCSF Tumor Immunology)

·         SDS-PAGE as the Second Dimension (UCSF Tumor Immunology)
In this method, proteins are highly denatured and associated with the anionic detergent SDS. Proteins run through the stacking gel, then stack at the membrane where the ion fronts compact them. Upon entering the separating gel, the proteins become separated with lower MW proteins running faster than higher MW proteins.
  

·         Analysis of Proteins using Small Format 2D Gel Electrophoresis (Dr Phillip Cash)
Detailed protocol on 2D protein electrophoresis. 

Protein Electrophoresis in Agarose Gel 

·         Protein Electrophoresis in Agarose Gels (FMC)
In certain circumstances, electrophoresis of proteins in agarose gels has distinct advantages when compared to polyacrylamide gels. This protocol describes procedures for gel casting, sample preparation, and protein staining and recovery.

Gel Staining

·         SDS PAGE Staining Protocol (LTI)
General method for silver stain and coomassie blue stain with recipes

·         Coomasie Blue Staining of Protein Gels (William H. Heidcamp)
Coomasie Brilliant Blue R 250 is the most commonly used staining procedure for the detection of proteins. It is the method of choice if SDS is used in the electrophoresis of proteins, and is sensitive for a range of 0.5 to 20 micrograms of protein.

·         Silver Staining of Protein Gels (William H. Heidcamp)

·         Staining of Polyacrylamide Gels (UCSF Tumor Immunology)

o        Coomassie G250 

o        Fast Stain (Zoion) 

o        Imidazole-Zinc
  

·         Silver staining of PAGE gels

·         Silver Staining of Acrylamide Gels (Waters Lab)

·         Silver Stain for SDS PAGE (Hahn Lab)

·         Silver staining of protein gels

·         Cupric Chloride Staining : for SDS-PAGE gels (Mike A Dyer)
This protocol is 2-3 times more sensitive than coomasie blue staining, it is much quicker, and the gels can be stored at 4° for many months without protein degredation.

·         Ethanol-based SDS-PAGE Coomassie Blue staining (NWFSC)

·         Direct Densitometry of Protein Gels (William H. Heidcamp)

Recipes

·         SDS-PAGE Recipes (The Cell Biology and Cytoskeleton Group, HMS)