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Methods:Bradford Assay

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Contents

Purpose

  • Determine total protein concentration of a sample

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Materials Needed

  • Standard curve using BSA
  • Sample of sufficient concentration
  • Bradford reagent (see Pierce or Bio-Radfor a commercial kit or the original paper by Bradford[1])
  • Spectrophotometer capable of reading at 595nm

Protocol

Links

User notes

  • Bradford assays measure TOTAL protein concentration (any contaminating protein will be measured along with your desired protein)
  • Bradford assays work for ug/mL concentrations between the ranges of 1 - >1000 ug/mL total protein
  • Bradford assays are used for proteins of greater than 3kDa
  • Bradford assays are generally sensitive to detergent, but a detergent-compatible kit is available from Bio-Rad

References

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  1. Bradford, MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72 248-54 PubMed EcoliWiki page