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Extra resources for Current Protocols in Protein Science

Sample text

These assays are performed in phosphate buffer. Do not attempt to perform them in Tris buffer since the enzyme is unstable in Tris and either no activity or a rapid decrease in activity may be observed. 2. 5 M ammonium sulfate (final concentration 50 mM) 120 µl 4 mM NADH (final concentration 160 µM) 50 µl enzyme preparation. 3. Cover the cuvette with Parafilm, mix the contents by inversion and incubate in the spectrophotometer for 3 min. 25 M 2-oxoglutarate (final concentration 5 mM), invert to mix and allow the reaction to proceed for 10 min in the spectrophotometer.

The Ultracel Amicon YM 30 or Ultracel PLTK membranes are appropriate substitutes. Purification of Glutamate Dehydrogenase from Liver and Brain Centriplus concentrators (Amicon) may be an alternative for concentrating the enzyme from volumes of less than 60 to 70 ml. , Centriplus 50, MWCO 50 kDa). The sample containing enzyme is retained by the membrane. The authors have found this procedure to be successful with several other dehydrogenases. 12 Supplement 29 Current Protocols in Protein Science Osmotic Dehydration Although ultrafiltration is a highly effective procedure for concentrating relatively large volumes, it may be less effective and convenient with small volumes and high protein concentrations.

The Ultracel Amicon YM 30 or Ultracel PLTK membranes are appropriate substitutes. Purification of Glutamate Dehydrogenase from Liver and Brain Centriplus concentrators (Amicon) may be an alternative for concentrating the enzyme from volumes of less than 60 to 70 ml. , Centriplus 50, MWCO 50 kDa). The sample containing enzyme is retained by the membrane. The authors have found this procedure to be successful with several other dehydrogenases. 12 Supplement 29 Current Protocols in Protein Science Osmotic Dehydration Although ultrafiltration is a highly effective procedure for concentrating relatively large volumes, it may be less effective and convenient with small volumes and high protein concentrations.

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Current Protocols in Protein Science


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