strange and branch like bands on SDS-PAGE
Posted 30 June 2011 - 06:15 AM
Posted 30 June 2011 - 08:48 AM
Hi pEXP23....welcome to bioforum....it would help if you can post an image of your stained gel as well as more details about your protocol....meanwhile, don't despair......
Hi, i have run many SDS-PAGES during working on my projects but recently I faced a problem.to check the purity of a recombinant peptide I am loading that after purification with Ni-NTA but after staing the gel all bands are duiffused and in so strange shape. There are no sharp bands and just bands like branches!I tried to prepare all buffers freshly and also I run at 80V for the first layer and 120 for the second but stil I couldnot overcome. could you please help me in this case?
- hobglobin, personal comment about my beauteous photo......
Posted 01 July 2011 - 10:29 PM
Posted 02 July 2011 - 02:35 PM
Protein samples should never be stored at fridge temperatures unless they are in a lyophilised state. Always store at a minimum of -20 deg C.
Posted 02 July 2011 - 10:32 PM
Posted 03 July 2011 - 07:22 AM
I would suggest:
Change to newly prepared /purchased APS. Use it fresh.
Use a multichannel pipette to load your acrylamide in between the glass when you make your gel. DO not load in one-side.
Use a lower voltage to avoid overheating, better you run your PAGE inside ice-box (I always do that).
..."best of our knowledge, as far as we know this had never been reported before, though I can't possible read all the published journals on earth, but by perform thorough search in google, the keywords did not match any documents"...
"what doesn't kill you, makes you stronger"---Goddess Casandra reminds me to be strong
"It's all just DNA. Do it."---phage434
Posted 03 July 2011 - 10:26 AM
Posted 19 August 2011 - 11:55 AM
Edited by Melvin J Noe Gonzalez, 19 August 2011 - 11:58 AM.