lsek

Hi kokoakash,

Size of the histone protein is very small. You can try to transfer the protein at shorter time, e.g. 10 to 20 min which will be more than sufficient. Some people double layer the PVDF as precaution.

One very important note, histones are quite basic (positively charged). Although residual SDS should be able to neutralize this and render it negatively charged during SDS-PAGE, the amount of SDS is not sufficient/diluted during the transfer step and hence reverting the histones back to positively charged. If that is the case, you will sometimes see a faint histone band or not at all. To circumvent this problem, reverse the position of the PVDF during transfer, or you can wedge the SDS-PAGE gel with PDVF at both sides of the gel for this purpose (one for capturing the histone at the cathode while the other is to trap the actin protein at the anode).


Useful reference: http://www.millipore...proteintransfer

Good luck.