help: c-fos probe
Sprankle at ciit.org
Sun Jan 28 21:44:05 EST 1996
In article <4e8sd1$dai at masala.cc.uh.edu>, lhe at dna.bchs.uh.edu (Lihua He) wrote:
> Is there any one have good experience using c-fos probe on Northern blot? I
> have problem with mine because it has high backgroud binding.
Have you tried increasing your hybridization temperature? I got very nice
hybridization to my northerns (poly A+ RNA from rat and mouse liver)
hybridizing at 55 deg. C., usually for 2 to 3 days (although I think
overnight worked also--it's been a while). Hyb was followed by brief room
temp washes with 2X SSC, 1 wash at 55 deg. C with 1X SSC/0.1% SDS for 30
min and 1 wash at 55 deg. C with 0.1X SSC/0.1% SDS. I should add that my
probes were labeled with 32-P and I was using nylon membranes.
Good luck--feel free to e-mail me if you have questions.
e-mail: sprankle at ciit.org
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