I've started doing zymography. I know that during gel casting the gelatin is somehow fixed in the gel, but, why it does not move during electrophoresis, since gelatin is also a protein?
It won't move, essentially because it is forming a gel within the acrylamide matrix.
The substrate (e.g. gelatin or casein) is *co-polymerized* within the polyacrylamide
matrix.
Michael Wendl already explained you the reason why the substrate (e.g. gelatin or casein) does not move during electrophoresis. Due to substrate copolymerization with acrylamide it loses electrophorectic mobility.
For further reading I am sharing a combined pdf with relevant literature.
Hope that helps.
As mentioned by colleagues, the process of copolymerization with acrylamide makes gelatine immobile within the electrophoresis milieu.
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