Does anyone know how to extract protein bands from Native gel to detect the product of the enzyme activity? I need the pellet from the band for further detection and to get rid of the gel.
First, do you really need to get rid of the gel ? If you have a chromogenic substrate that can diffuse into the gel, you could perform a zymogram stain. If you do need to get the enzyme out of the gel, you may try to put the gel slice into a dialysis bag with electrophoresis buffer and put the bag across a horizontal gel tank (like those used for DNA agarose gels) and taking care that the gel piece faces the negative electrode (I assume that your protein is negatively charged). Run the current at the highest voltage that doesn't heat up the buffer for 1-2 h, then invert voltage for 1 min. Collect the content of the bag, rinse bag with some buffer, followed by concentration of the eluate in a centrifugal concentrator (Vivaspin, Centricon or the like).
Thank you very much for your suggestion. My aim is to get out the product of the enzyme (not the enzyme), the stained product is lead sulfide (PbS) which i need to prove it after digestion. Is it possible to do?
Usually activity gels (zymograms) are run under native conditions and detection of target proteases is carried out by using specific substrates against target proteases. It is not clear to me what is the reason to detect the product of the enzymatic activity in gel. You could electroelute the required band from gel and run enzymatic assays if protein is still active.