Author Topic: Problem to transfer small proteins  (Read 3084 times)

GE Maria

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Problem to transfer small proteins
« on: November 18, 2011, 03:39:16 AM »
Lately I had a college that experienced poor transfer of small proteins, what can be done here?

This could be a problem if you wish to transfer both large and small proteins at the same time, by using lower voltage or shorter transfer time the large proteins will not be efficient transferred. Here I would suggest that you place two membranes to each other so if the small protein will migrate through the first membrane it will be cached on the second membrane.

More generally if you have problems to transfer small proteins consider this!
Optimize transfer time using relevant molecular markers.

Use a PDVF membrane, which has a higher protein binding capacity than nitrocellulose membranes

Use membranes with optimal pore size

Use a higher percentage or gradient gel to help retain proteins during equilibration and transfer

Traces of SDS in the gel may interfere with the binding of small proteins to the membrane, so equilibrate the gel in transfer buffer for at least 15min  in the transfer buffer to remove the SDS. Little higher methanol concentration in the transfer buffer (15-20%) can help to remove the SDS.

Hope this can be of some help!
Maria