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.

Contents

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 -
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