Detection of proteins on blot transfer membranes.
In the basic and alternate protocols of this unit, proteins are stained after electroblotting from polyacrylamide gels to blot transfer membranes. If the samples of interest are electrophoresed in duplicate and transferred to a blot transfer membrane, half of the membrane can be stained to determine the efficiency of transfer to the membrane and the other half can be used for immunoblotting (i.e., western blotting). Detection limits of each staining method are given along with a list of compatible blot transfer membranes and gels. A support protocol describes a method for alkali treatment that enhances subsequent staining of bound proteins.
Sasse
Joachim
J
Shriners Hospital for Crippled Children, Tampa, Florida, USA.
Gallagher
Sean R
SR
D002244:Carbon; D004396:Coloring Agents; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D005456:Fluorescent Dyes / Q000032:analysis; D017979:Gold Colloid; D006878:Hydroxides; D007202:Indicators and Reagents; D008567:Membranes, Artificial; D009942:Organometallic Compounds; D010781:Photography / Q000379:methods; D017680:Potassium Compounds; D011506:Proteins / Q000032:analysis*; D013194:Staining and Labeling / Q000379:methods*
D004396:Coloring Agents; D005456:Fluorescent Dyes; D017979:Gold Colloid; D006878:Hydroxides; D007202:Indicators and Reagents; D008567:Membranes, Artificial; D009942:Organometallic Compounds; D017680:Potassium Compounds; D011506:Proteins; C515601:Sypro Ruby; C028433:chinese ink; C058457:Aurodye; D002244:Carbon; C029943:potassium hydroxide