Lab Manual
biological reagents
Cell Free Systems
In Vivo Systems
misc methods

Purification of Proteins from Inclusion bodies.

Two protocols have been applied in our lab to purify proteins from inclusion bodies: Sarkosyl Method or Lin-Cheng Method.

Sarkosyl Method:

In this method cells lysed by sonication in the presence of lysosyme. After centrifugation pelleted protein (inclusion bodies) is solublized with 1 % sarkosyl.

STE Buffer 100 mM NaCl
10 mM Tris-HCl pH 8.0
STE-S STE Buffer (1 % Sarkosyl)
STE-T STE Buffer (1 % Triton X-100)


This procedure is written for a 150 mL culture you may scale it up or down accordingly.


1. Cells from 150 mL culture were harvested in the Beckmann J2-21 in a JA-20 rotor at 5000 rpm for 15 minutes @ 4C.

2. Resuspend the resulting pellet in 3 mL of STE buffer.

3. Add lysozyme to a final concentration of 150 g/mL and place on ice for 30 minutes.

4. Sonicate on ice, using a 40 second sonication and a 20 second rest period (set to 40 and use the small probe). Repeat an additional three times.

5. Aliquot into two 1.5 mL microfuge tubes.

6. Spin for 5 minutes @ 4C on maximum. (Keep the supernatant for further analysis).

7. Resuspend the pellet in 0.1 mL of STE-S. This will require vigorous pipetting (the pellet may not be completely soluble).

8. Centrifuge for 2 minutes at 4C and transfer the supernatant to a 1.5 mL microfuge tube.

9. Add 9 volumes of STE-T to the supernatant (to form a mixed-micelle with sarkosyl).

10. Samples can be frozen on dry ice or liquid nitorogen and stored in the -80C freezer until required.