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protein electrophoresis SDS-page
#1
1. I run protein SDS-page. The beginning, the protein bands were appearance, but then running back, i did not see any band. Who answer me why? I asked my teacher, he said that contents of protein not enough. But after condensed sample that shows no band.
2. I run SDS-page according to the method of Laemmli (1970), I mixed stacking gel 3%, it was polymerized about 15 minutes . But after I also do the same protocol that was not polymerize to 1 hour. Who can explain to me?
Here are recipes stacking gel 3% of Laemmli protocol:
2.5 ml 4X Stacking Gel Buffer
6.5 ml dH2O
1.0 ml acrylamide (29: 1)
200 microliter of 10% APS
20 microliter of TEMED
* 4X Stacking Gel Buffer
0.5 M Tris pH 6.8 6,06g, pH to 6.8 with HCl
0.4% SDS 0.4 g SDS
up to 100 ml with dH2O
3. Protein of salted squid less protein of fresh squid. Is protein of salted squid went outside in salted environment?
Thank you very much!
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