native or sds, acrylamide should polymerize in presence of APS. I usually use 10ul of temed and 100ul of APS, for 50ml of SDS-PAGE solution. the excess free radicals may lead to protein degradation or altered enzyme activity
10% aps is best for good polymerization.more % of aps will not affect your gel concentration but sometimes cause negative impact on the protein separation.
How do you store your APS? APS will easily traps moisture. It will reduce performance, thus makes your gel takes longer time to polymerize. 10 % of APS works best in my opinion.
thank u for suggestions.But i have problem that the temperature is not constant so the time period for polymerization requires 3-4hrs if i will take 10% APS. next query is that What is the time period for polymerization? Is their specific time or not?
Temp range of 20-45 deg should not affect the polymerization. You can prepare the gel late in the evening, cover it with plastic film a day prior to your experiments.
I agree with Wajeeha Asad, check if your products are not expired. it is not necessary to rise the % of APS.
If you have a problem of temperature in your room (too low for example) you could accelerate the polymerization by placing the gel in front of a lamp. The heat of the lamp will accelerate the polymerization (I made it in winter when temperature in lab down and my gels made sevarl hours to polymerize...)