Can buffer affect banding patterns on SDS-Page gels?

2 posts / 0 new
Last post
hkamrud
hkamrud's picture
Can buffer affect banding patterns on SDS-Page gels?

I am analyzing the autolysis of bovine trypsin and have run a gel with the trypsin samples varying in temperature and time.  Trypsin is about 23 kiloDaltons and I can identify that band, but I have a band that shows up with just the trypsin and buffer, and also with the trypsin, buffer, and inhibitor (the inhibitor has a molecular weight of about 28 kD). the band is actually much darker in the inhibitor samples even though the concentration of trypsin protein is the same.  The trypsin sample was given to me in a tris buffer (MW 121 kD) and I used a Phosphate buffered saline.  Could the buffers I'm using affect the banding pattern?

g a
g a's picture
Dear Hkamrud

Dear Hkamrud

Although I dont have the direct answer to your question but perhaps this may help.
Running buffer does have influence on the baning pattern...... for instance, using glycine as running buffer and 10%gel resolution limit remains closer to 20kDa. using tricine the same % of gel can resolve 1000da peptides as well.
and even if yre using the glycine as running buffer, but using a resolving gel at pH 9.2 (instead of usual 8.8) the same resolution can be achieved.

Bottom line: Buffer does play critical roles in electrophoresis.