Bio-Rad ReadyPrep Protein Extraction Kit (Signal) User Manual
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Section 4
Instructions for Use
Note: Chill buffers S1 and S2 on ice for at least 15 min
before beginning (invert bottles several times during the
incubation).
1. In a microcentrifuge tube, add 0.5 ml of buffer S1 per
25–50 mg of animal tissue or 0.05 ml of wet cell
pellet from sources such as cell culture, yeast, or
bacteria. For plant tissue add 0.5 ml of buffer S1 per
0.25 g tissue. The sample-to-buffer volume ratio
indicated above is only a guide and may be adjusted
depending upon the desired scale of the preparation.
Notes: Protease inhibitors may be added to Buffer S1
immediately prior to use to prevent proteolysis during
extraction.
Insufficient volume of buffer S1 may result in poor cell lysis
and contamination of the signal protein fraction with
cytoplasmic protei ns.
Plant tissue should be ground to a fine powder using a
mortar and pestle in liquid nitrogen before addition of
buffer S1.
2. Sonicate the suspension on ice with an ultrasonic
probe to break open the cells and fragment the
genomic DNA. During sonication, care must be
exercised to prevent heating of the sample. Sonicate
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