Dear Oldcloner Thank you for your suggestion.
I did try the use my protein extraction with my lab mates buffer (it is a comercial buffer) and the results looked similar to mine so I assumed that my protein have a problem.
my sample is Hacat cell, I did not count the cell for protein extraction I just cultured them in the 60 mm dish and harvest it and did the protein extraction.
I would try as your suggestion using my sample with my lab mate's gel and my gel with my lab's protein and let's see the results.
Dear mdfenko thank you so much for your question?
I followed the protein lysis buffer product extraction protocol, I used bradford protein assay. How to know if my protein has been succesffuly hydrolyzed?
I thought I prepared buffers properly, I would try to make a new buffer again
tracking gel traveled through the end of gel
sample color was clear bue after adding the sample buffer
gel component is 15% gel (H2O 2.3 ml; 30% acrylamide mix 5 ml; 1.5 Tris (pH 8.8) 2.5 ml; 10% SDS 0.1 ml; 10% ammonium persulfate 0.1 ml; TEMED 0.004 ml) and 5% stacking gel.
running condition is 60V for 30 min and then 100V for 90 min.
on the right is the standard size (protein marker) I use 2 microlite.
please give me any suggestion if I do something wrong.
I would try as OldCloner suggest first.