Search found 6 matches

by mutant11
Tue Nov 22, 2011 2:39 am
Forum: Molecular Biology
Topic: Western Blot TroubleShooting
Replies: 7
Views: 6792

Thanks for the advice Daniel!
by mutant11
Mon Nov 21, 2011 5:08 am
Forum: Molecular Biology
Topic: Western Blot TroubleShooting
Replies: 7
Views: 6792

I am not able to detect endogenous levels of the transcription factor. I was wondering if you had any suggestions on improving the signal of my protein. Would it be worth it to do a nuclear lysate?
by mutant11
Mon Nov 21, 2011 3:11 am
Forum: Molecular Biology
Topic: Western Blot TroubleShooting
Replies: 7
Views: 6792

Re: Western Blot TroubleShooting

I harvest my samples using RIPA buffer + protease inhibitor on ice. I use a 6 well dish and I scrape the samples off on ice. I rotate the samples at 4C for 1 hour for lysis. I then centrifuge for 30 min. at 13,000 rpm at 4C. I take off the supernatant. I add 2X SDS PAGE loading dye and heat at 100C ...
by mutant11
Fri Nov 18, 2011 10:17 pm
Forum: Molecular Biology
Topic: Western Blot TroubleShooting
Replies: 7
Views: 6792

Western Blot TroubleShooting

Dear all, I need some help with Western Blots. I am trying to detect a transcription factor that is expressed at pretty low levels endogenously in cell lines. Does anyone have any suggestions? -I use RIPA buffer with protease inhibitor to harvest the protein. I keep everything on ice. -I incubate in...
by mutant11
Mon Jul 04, 2011 11:55 pm
Forum: Cell Biology
Topic: Growing MDCK Cells on Coverslips
Replies: 1
Views: 2818

Growing MDCK Cells on Coverslips

Hi all, Does anyone work with MDCK cells? I want to seed them onto glass coverslips and have been using glass coverslips coated with poly-L-lysine. The cells sometimes do not stick well to the coverslips and I have found that the transient transfection rate is lower on the coated glass slides than j...
by mutant11
Sat May 14, 2011 7:44 pm
Forum: PDF & Doc Files
Topic: URGENT: Need a paper
Replies: 2
Views: 10532

URGENT: Need a paper

Hi, I need to read this paper for my upcoming qualifying exam and I can't find it anywhere! E.S. Kim, M.S. Kim and A. Moon, TGF-beta-induced upregulation of MMP-2 and MMP-9 depends on p38 MAPK, but not ERK signaling in MCF10A human breast epithelial cells, Int. J. Oncol. 25 (2004), pp. 1375–1382. My...