I just need an assistance about how to develop a blot , using luminata forte and Xray cassete? Like the conditions in which the developer, fixer or the luminata forte to be opened... Is complete darkness mandatory?
in short, the procedure to be followed after a successful transfer, primary/ secondary antibody probing, as i am facing some problems while developing my blot, like the xray film gets exposed even after being opened in dark and my target protein band develops very thin despite normal loading control development...
hi
1- you should work in fully dark room with red dark room lamp ( such as Philips brand) X-ray film not sensetive for RED light.
2- Luminate forte 5 min in PVDF and then semi-dry your PVDF and then put your PVDF in nylon sheet .
3- expose time start from 30 sec to 20 min . routinely 5 min is ok
good luck
Thanks Reza
Your suggestion is fine. However, i want to know whether luminata forte can be applied to the PVDF membrane in white light? or luminata forte is light sensitive which needs to be opened in dark room with red lamp?
hi
luminata forte not sensitive to whit light . just X-Ray Film sensitive to wight light.
feel free
Thanks Reza
It means i can pour luminata forte to the membrane in white light, then i will put off the light and develop the blot, using x-ray film in red lamp....
Thanks again
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