I AM USING 20% METHANOL IN TRIS GLYCINE , 0.2 MICRON
NITROCELLULOSE MEMBRANE FOR 100 V HR . the ponceau staining looks weaker compared to precast gel 10-12% with the transfer using sds and ethanol.
please suggest me the exact time to transfer histone H4
Divaker Choubey
If possible, I would use PVDF membrane. If not possible, use 100 mAmp for 2-2.5 h. Not many major proteins are detected in the small size range. Immunoblotting using anti-H4 is a better method to assess the transfer conditions.
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Julie Joseph
In my experience, 1-1.5 hours is sufficient. Make sure your transfer is done on ice.
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Abhishek Jauhari
Use Wet transfer for 125 minutes at 250 mA current.
Make sure your transfer buffer should be chilled or transfer in cold room or under 4 degree refrigerator.
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