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Reducing Agents

Reducing agents are responsible for the reduction of disulfide bonds within proteins and peptides.  TCEP HCl, DTT and BME have been shown to reduce protein aggregation that may inhibit crystallization.

Reducing agents can interact with metals within your sample resulting in poor efficiency of your reducing agent.  If possible it is suggested to use a metal chelating agent such as EDTA to remove all undesired metals.  Unfortunately, this may not be possible since many enzymes require metal in order to properly function. BME is especially sensitive to copper, cobalt and many phosphate buffers. DTT is sensitive to nickel therefore caution should be used when if used in combination with affinity chromatography.

Relative Strength: TCEP > DTT > BME

Reducing Agent Summary Table:

  Name   Solubility in H2O   R.T. Exposure   pH Range   Odor   Applications
  TCEP HCl   310 g/L   2-3 weeks   1.5-11.1   No   Mass spectrometry
  DTT   50 g/L   3-7 days   ~7   Yes   DNA Deprotecting Agent
  BME   1:1 Ratio   2-3 days   ~7 (5.5-10)   Yes   SDS-PAGE

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