Can anyone suggest how to optimize resolution for plant seed proteins.

05 April 2013 1 1K Report

7 CM IPG Strip with 100 microgram of protein, 8 M urea, 2 M Thiourea, 2% CHAPS, 0.5% Ampholytes (Biorad).Also it take more than eight hrs for focusing to complete.

Step 1 250 V for 20 min, Step 2 4000 V (Lenear), Step 3 4000 V (Rapid), 10,00 V Hours

Prafull Gadge

Dear Shankar here are answers to your questions:

Extraction of seed proteins was carried out in distilled water without any buffer followed by acetone precipitation.

I want to carry out Zymography thus to keep proteins in non denaturing conditions DTT was not added.

Passive re hydration was carried out for 12 hours.

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