蛋白质技术

Western Blotting Using the SemiPhor Semi-Dry Transfer Unit Remove the stacking gel from the gel. Measure the gel and record its size. Do not pre-soak the gel. Do not cut a corner off the gel as this m ...

Studying Protein Interactions by Far-Western BlottingFar-Western blotting was originally developed to screen protein expression libraries with 32P-labeled glutathione S-transferase (GST)-fusion protei ...

Too many bands on a Western blot Western- or immunoblotting is a commonly employed technique for the detection of protein antigens in complex mixtures. Samples are first separated by SDS-polyacrylamid ...

Too many bands on a Western blot Western- or immunoblotting is a commonly employed technique for the detection of protein antigens in complex mixtures. Samples are first separated by SDS-polyacrylamid ...

1. Perform SDS-polyacrylamide gel electrophoresis (SDS-PAGE) on a cell sample prepared using modified RIPA buffer and transfer the proteins to a nitrocellulose membrane. 2. Wash the nitrocellulose t ...

1. Perform SDS-polyacrylamide gel electrophoresis (SDS-PAGE) on a cell sample prepared using modified RIPA buffer and transfer the proteins to a nitrocellulose membrane. 2. Wash the nitrocellulose t ...

Troubleshooting Guide: Western BlotProblem: Poor TransferPossible SourceTest or ActionMembrane does not wet uniformlyPre-wet membrane in 100% methanolMW of protein is less than 10000If protein has MW ...

Preparation of Brain Cytosol1. Chill a glass plate on ice. 2. Place brain tissue on the chilled glass plate and cut slices of brain into small pieces. 3. Resuspend the pieces in ice-cold RIPA buffer w ...

Protease Inhibitors from Alberts' Lab Protocols (Kathy Miller/Chris Field)Protease inhibitors are rapidly inactivated in aqueous solutions and should be added to solutions immediately before use. Cock ...

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 ...