Hi,

1. Proteins separated on polyacrylamide gels can be transferred onto immobilized membranes (blotting). However, in many instances, a gentle method to elute the proteins directly into a liquid phase is required to avoid chemical modification or denaturation of the eluted proteins. Common approaches include elution by diffusion and electroelution (Deutscher, 1990; Waterborg and Matthews,1994).

2. In the diffusion method, the protein band or spot of interest is sliced from the gel and minced into small gel pieces to increase the diffusion surface. These gel pieces are then mixed in a SDS solution with gentle agitation until the protein fraction diffuses out of the gel (Deutscher, 1990).

Try these protocols and go through the references what am given. It will helpful to you.

All d Best

Hi,

1. Proteins separated on polyacrylamide gels can be transferred onto immobilized membranes (blotting). However, in many instances, a gentle method to elute the proteins directly into a liquid phase is required to avoid chemical modification or denaturation of the eluted proteins. Common approaches include elution by diffusion and electroelution (Deutscher, 1990; Waterborg and Matthews,1994).

2. In the diffusion method, the protein band or spot of interest is sliced from the gel and minced into small gel pieces to increase the diffusion surface. These gel pieces are then mixed in a SDS solution with gentle agitation until the protein fraction diffuses out of the gel (Deutscher, 1990).

Try these protocols and go through the references what am given. It will helpful to you.

All d Best

You have to slice the spots of interest, and cut it .. I use to cut pipette tip.

Then you must make a reduction of gel and Spots of interest will be subjected to tryptic in-gel digestion.

http://link.springer.com/protocol/10.1007%2F978-1-61779-821-4_33

http://www.pall.com/main/laboratory/literature-library-details.page?id=2273

I hope, these links will help you

Hi, I go with praveenkumar , here i attach a procedure that you may follow

Thanku