SDS - PAGE for Identifying nonmucin protein in BSM
Prepare the sample solutions :
prepare the mucin samples .
Assign different volume for each sample , 10ul , 20ul , 30ul , respectively .
Add DDI water to the samples , let each tube has 30ul solution .
Add loading buffer to each sample .
Heating them in 95 degree for 10min .
Loading the running buffer .
Add some running buffer into the electrophrosis cell .
peel the green paste at the bottom and take away the top green plastic cover , set the gel in the electrophorsis cell .
Fill the inner and outer buffer chambers with running buffer .
Make sure that the level of the running buffer above the top of the gel and there is no leakage from the inner buffer chamber to the outer .
Performing electrophorsis .
Load the protein sample on the gel .
Connect the electrophoresis cell to the power supply and perform electrophores in Run conditions : 150 V ~ 200 V
Run time : until the loading buffer line come to the bottom of the gel Staining .
Wash gels in DI H2O .
Remove all water from staining container and add stain ( PageBlue ) , agitate overnight .
Wash gel using DI water , first time 5 minutes ; second time 10 minutes ; third time 30 minutes ; forth time , 1 hour and fifth time keep the gel in DI water agitate overnight .
