Table 1. Lysis buffers used for total proteome extraction from B. anthracis spore suspension.
|
№ |
Reagent |
Final concentration in solution |
Amount per 100 ml solution |
|
1 |
Tris basic |
40mM |
0.4846 g |
|
2 |
DTT |
10mM |
0.15425 g |
|
3 |
Triton X-100 |
2% |
2 ml |
|
4 |
Glycerol |
10% |
10 ml |
|
5* |
Bacterial Protease Inhibitor Cocktail |
1 μg / ml |
6 μl |
|
6 |
Guanidine chloride |
6М |
70.856 g |
|
7 |
Urea |
8М |
48.048 g |
|
8 |
Thiourea |
2М |
15.224 g |
Note. Explanations for table 1: components 1-5 are used in the preparation of both buffers, item 6 is used in the preparation of lysis buffer with 6M guanidine chloride, items 7 and 8 - lysis buffer with 8M urea / 2M thiourea.
* Bacterial protease inhibitor cocktail is added immediately before the start of the work. The pH of the lysis buffer with urea / thiourea is 8.5-9.0. The pH of the lysis buffer with guanidine thiocyanate is 7.5-8.0.