Barry Canton/Bacterial lysis
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I need to lyse some E. coli under denaturing conditions. I'm collecting my on how to best do that here.
OWW protocols
- Sauer:Purification of His-tagged proteins/Denaturing prep - the lysis part of this should generally useful
- Knight:Purification of His-tagged proteins/Denaturing - slightly different lysis buffer
- RNA extraction - describes use of guanidine isothiocyanate
- Sauer:ClpX purification/Protocol 1
- Sauer:His6 ClpX purification
- Sauer:SspB purification - Tween
- Sauer:Lysing E. coli with Lysozymes - lysozyme methods for native purification
- Sauer:Expression/purification of 35S-Met proteins/Native purification - Imidazole
- WangLab:Lysis Buffer
Chemical lysis
Lysing reagents
- NaH2PO4, Guanidine hydrochloride, imidazole
- Urea, TCEP, imidazole, NaH2PO4
- Lysozyme and guanidine isothiocyanate
- DTT, PMSF
- NaH2PO4, imidazole, DTT
- beta-ME, Tween-20, PMSF
- NaH2PO4, Imidazole
- Triton, SDS, NAF
Chemical functions
- Guanidine hydrochloride - denatures protein
- NaH2PO4 - pH buffer
- Imidazole - used for his-tagged protein purification
- Urea - denatures protein
- TCEP - breaks disulfide bonds
- Guanidine isothiocyanate - denatures protein
- DTT - breaks disulfide bond
- beta-ME - breaks disulfide bonds
- Tween - detergent
- PMSF - serine protease inhibitor
- Triton - nonionic detergent
- SDS - detergent
- NAF -