Hello,
Recently I had an issue with polymerization of SDS page. I used to add 5 ul TEMED and 50 ul of 10% APS for each 12 ml of SDS page but suddenly my SDS page stopped polymerizing and could take hours to solidify. So, I increased TEMED to 10 ul and APS to 100 ul and the polymerization went very well. However, I am getting messy protein separation since after with so many unexpected bands that may look like the protein is degraded. Does this have anything to do with increasing APS/TEMED?
THANKS
hello.
I don't think that APS or TEMED would influence to protein separation, but you can check the expiration date of the agents.
Is your target protein a complex? the subunit can be dissociated while you prepare the samples for the SDS-PAGE.
Hi there,
Protein degradation when catalyzed by proteases always occurs prior SDS/PAGE. I don't think gel reagents affect proteins... Any picture of the before/after comparison?
I'd just get some fresh APS, its quite unstable and unless you plan to publish on the impact of old APS on protein stability I'd move on! Good luck!
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