蛋白组学

Abstract: Many people have vented out frustration over insoluble GST-fused proteins. This is a protocol for enzymatically active soluble GST-fused proteins. All GST-fused proteins are rendered soluble ...

Troubleshooting Tips for Western ImmunoblottingThe following tips can be used to overcome the most common problems encountered during Western blotting. Smeared Pattern or Distorted Bands Uneven conta ...

Troubleshooting Tips for Western ImmunoblottingThe following tips can be used to overcome the most common problems encountered during Western blotting. Smeared Pattern or Distorted Bands Uneven conta ...

Bacterial Protein Extraction - mini scale - SonicationProtein Extraction 1) Resuspend pellet of 10ml cell culture in 1ml lysis buffer (or 100ml bacterial culture for very low expression level). Sugges ...

Preparation of cell lysates from E. coli by enzymatic lysisMaterialsChemicalslysosymeLysis buffer50 mM Tris-HCl pH 7.5 50-200 mM NaCundefined5% glycerol (v/v)1 mM DTT1 mM PMSF ...

Sonication of BacteriaThis procedure is used to break cells that contain a protein to be purified. This procedure is valid only for the use on Appling's lab sonicatior. PREPARATIONPlace your sample in ...

Preparation of cell lysates from E. coli by sonicationMaterialsLysis buffer50 mM Tris-HCl pH 7.5 50-200 mM NaCundefined5 mM DTT1 mM PMSF ...

Preparation of cell lysates from E. coli by sonicationMaterialsLysis buffer50 mM Tris-HCl pH 7.5 50-200 mM NaCundefined5 mM DTT1 mM PMSF ...

Preparation of cell lysates from E. coli using a French PressMaterialsLysis buffer 50 mM Tris-HCl pH 7.5 50-200 mM NaCundefined1 mM MgCl25 mM DTT1 mM PMSF ...

Preparation of cell lysates from E. coli using a French PressMaterialsLysis buffer 50 mM Tris-HCl pH 7.5 50-200 mM NaCundefined1 mM MgCl25 mM DTT1 mM PMSF ...

Preparation of cell lysates from yeast by enzymatic digestion of the cell wallMaterialsChemicalsLyticase USA)Brij 58Working buffer50 mM Tris-HCl pH 8.0 10% sorbitol ...

Preparation of cell lysates from yeast by enzymatic digestion of the cell wallMaterialsChemicalsLyticase USA)Brij 58Working buffer50 mM Tris-HCl pH 8.0 10% sorbitol ...

Preparation of cell lysates from yeast using a French PressEquipment and reagentsLysis buffer50 mM Tris-HCl pH 7.5 50-200 mM NaCundefined1 mM EDTA 5 mM DTT1 mM PMSF1 µg/ml leupeptin1 µg/ml pepstatin A ...

Preparation of cell lysates from yeast using a French PressEquipment and reagentsLysis buffer50 mM Tris-HCl pH 7.5 50-200 mM NaCundefined1 mM EDTA 5 mM DTT1 mM PMSF1 µg/ml leupeptin1 µg/ml pepstatin A ...

Solubilization and renaturation of proteins from inclusion bodiesMaterialsWash buffer50 mM Tris-HCl pH 7.5 50-200 mM NaCl ...

Preparation of cell lysates from yeast using glass beads vortexingEquipment and reagentsLysis buffer50 mM Tris-HCl pH 8.0 1% DMSO50-200 mM NaCundefined1 mM EDTA1 mM PMSF1 µg/ml leupeptin1 µg/ml pepstatin A ...

Solubilization and renaturation of proteins from inclusion bodiesMaterialsWash buffer50 mM Tris-HCl pH 7.5 50-200 mM NaCl ...

Large scale nuclear extract preparationHattoti's protocol adapted to cell culture:Hattori M Tugores A Veloz L Karin M Brenner D (1990) A simplified method for the preparation of transcrip-tionally act ...

Solutions:Buffer A (Hypotonic Buffer): 1L10 mM HEPES pH 7.9 10 ml 1M HEPES pH 7.91.5 mM MgCl2 1.5 ml 1M MgCl210 mM KCl 3.33 ml 3M KCl0.5 mM DTT 500 µl 1M DTT0.2 mM PMSF 1 ml 0.2 M PMSFBuffer B (S100 e ...

Preparation of Nuclear Extracts for Gel Shifts and Westerns Blots ...