Bio-Rad ReadyPrep™ Protein Extraction Kit (Membrane II) - Manual (Page 10)

Section 5
Instructions for Use
Extraction of 100–200 mg of sample.
1. Immediately before performing an extraction, prepare
Lysis Buffer, Membrane Protein Concentrating Reagent
and ReadyPrep 2-D rehydration/sample buffer 1 per
the instructions in Section 4. For a standard extraction
you will need approximately 15 ml of Lysis Buffer,
60 ml of Membrane Protein Concentrating Reagent
and 1 ml of 2-D Rehydration/Sample Buffer 1. Chill
both the Lysis Buffer and the Membrane Protein
Concentrating Reagent on ice for 10–15 min before
proceeding.
2. In a 2.0 ml microcentrifuge tube (on ice), add 1 ml of
Lysis Buffer per 100–200 mg of animal tissue or wet
cell pellet from sources such as cell culture, yeast, or
bacteria. For plant tissue add 2–3 ml of Lysis Buffer
per each gram of tissue in, for example, a disposable
14 ml round-bottom tube. The sample-to-buffer
volume ratio indicated above is only a guide and may
be adjusted depending upon the desired scale of the
preparation and type of sample used.
Insufficient volume of Lysis Buffer may result in poor cell
lysis and incomplete solubilization of proteins.
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