Christopher Wilson

Questions and Answers (6) View all

• Answer added in Affinity Chromatography
  8 Problem in re-binding of maltose binding protein to amylose resin
  By Muhammad Adnan · University of Leicester
  Christopher Wilson · Brandeis University

  Yes this is common for me. MBP not binding after removal of the fusion protein happens in about 50% of my constructs and seems to be construct specfi... [more]

  Yes this is common for me. MBP not binding after removal of the fusion protein happens in about 50% of my constructs and seems to be construct specfic. The best thing to do is to stick a His tag on the N-terminal of MBP. and pass over a nickel or cobalt column after cleavage. This works 100% of the time.
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• Answer added in Expression Profiling
  41 Irreproducible protein expression
  By Mark van Raaij · Spanish National Research Council
  Christopher Wilson · Brandeis University

  See Meena Ali's response. Is the insert codon optimized? + this is a common feature in a lot of proteins. where some colonies produce more protein ... [more]

  See Meena Ali's response. Is the insert codon optimized? + this is a common feature in a lot of proteins. where some colonies produce more protein then others. One thing you could do is test a number of different colonies and make a glycerol stock from a colony you know expresses the protein well.
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• Answer added in Enzymology
  12 Reducing agent to break disulfide bonds in peroxidase enzyme: TCep or dtt?
  By Khawar Siddiqui · University of New South Wales
  Christopher Wilson · Brandeis University

  You must use TCEP. DTT loses its reducing power at a ph lower then 7.0. Also DTT breaks down rapidly while TCEP is stable. 

  You must use TCEP. DTT loses its reducing power at a ph lower then 7.0. Also DTT breaks down rapidly while TCEP is stable.
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• Answer added in Protein Purification
  25 A problem of "disappearing" protein during concentration of the sample
  By Michal Zdzalik · Jagiellonian University
  Christopher Wilson · Brandeis University

  Just because you can't see the protein percepitate doesn't mean that its not aggregating. Lots of proteins form high weight soluble aggregates with d... [more]

  Just because you can't see the protein percepitate doesn't mean that its not aggregating. Lots of proteins form high weight soluble aggregates with different spectral properties then free protein. You can check for prescence of soluble aggregates using dynamic light scattering.
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• Question asked in Simulation and Modeling
  2 Aligning distant structures using a common ligand.
  Does anyone know a quick way to align a group of structures, say 30, through a common ligand found in the structures? The idea is that the structures... [more]
  Does anyone know a quick way to align a group of structures, say 30, through a common ligand found in the structures? The idea is that the structures are distantly related but all have this common ligand bound.
  By Christopher Wilson · Brandeis University
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