The question is about how strongly the proteins bind to the membrane. Is the rubbing of my hand going to significantly affect the retention of the proteins on the membrane? Not that I will be doing such a thing, but want to know.
Every effort should be made to avoid abrasion of the membrane surface, which can smear the bands, strip off the blocking agent, and cause other artifacts. The membrane should preferably not be handled except at the edges where nothing of importance is bound. Once the membrane is in a container after the transfer, there is no need to touch it at all until it is transferred to an imaging device.
Hi Jatin,
Since you are transferring by electroblotting the protein onto the membrane, it is not possible to remove the protein from the membrane by physical rubbing. However, hands contain skin proteins and you really should try and not touch the proteins using bare hands. If you are using "powder free" gloves, try to touch the 4 corners of the membrane where there are no proteins transferred.
Good luck,
Hediye
Hello Adam and Hediye,
I understand that smearing, contamination, and non-specific binding (if blocking agent is removed) can be a problem. If i got it right, the proteins will still not come out of the membrane totally.
Thank you for the help.
Regards,
Jatin
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