Barry Canton/Bacterial lysis

I need to lyse some E. coli under denaturing conditions. I'm collecting my on how to best do that here.

OWW protocols

1. Sauer:Purification of His-tagged proteins/Denaturing prep - the lysis part of this should generally useful
2. Knight:Purification of His-tagged proteins/Denaturing - slightly different lysis buffer
3. RNA extraction - describes use of guanidine isothiocyanate
4. Sauer:ClpX purification/Protocol 1
5. Sauer:His6 ClpX purification
6. Sauer:SspB purification - Tween
7. Sauer:Lysing E. coli with Lysozymes - lysozyme methods for native purification
8. Sauer:Expression/purification of 35S-Met proteins/Native purification - Imidazole
9. WangLab:Lysis Buffer

Chemical lysis

Lysing reagents

1. NaH2PO4, Guanidine hydrochloride, imidazole
2. Urea, TCEP, imidazole, NaH2PO4
3. Lysozyme and guanidine isothiocyanate
4. DTT, PMSF
5. NaH2PO4, imidazole, DTT
6. beta-ME, Tween-20, PMSF
7. NaH2PO4, Imidazole
8. Triton, SDS, NAF

Chemical functions

1. Guanidine hydrochloride - denatures protein
2. NaH2PO4 - pH buffer
3. Imidazole - used for his-tagged protein purification
4. Urea - denatures protein
5. TCEP - breaks disulfide bonds
6. Guanidine isothiocyanate - denatures protein
7. DTT - breaks disulfide bond
8. beta-ME - breaks disulfide bonds
9. Tween - detergent
10. PMSF - serine protease inhibitor
11. Triton - nonionic detergent
12. SDS - detergent
13. NAF -