Biotech lab 3-staining SDS-PA gels-background Flashcards
1
Q
What stain will we be using?
A
Coomasie Brilliant blue
2
Q
What is coomasie brilliant blue?
A
Aminotriarylmethane dye which forms strong complexes and stoichiometrically binds with proteins but does not bind tightly to the SDS-PAGE gel.
3
Q
How are proteins separated by SDS-PAGE fixed?
A
Using either methanol or ethanol:glacial acetic acid then stained with coomasie blue.
4
Q
How long is the gel immersed in the fixing solution?
A
30 minutes
5
Q
The uptake of the dye is proportional to what?
A
The amount of protein.
6
Q
How is un-incorporated dye allowed to diffuse?
A
In a de-staining step.
