Bio 499CB (Loomis Cancer)

Kari Loomis honors thesis seminar

Librarian does workshop on library research.

We do Bradford assay lab:

Explain Bradford assay.

Add some Bradford reagent to some BSA - Ask how much protein?

Need standards to compare to!

• Bradford concentrate from Bio-Rad 500-0006
• dilute 1:4 with water
• Make BSA solution 0.1 mg/mL (dilute from the test 2 m/mL test solution
• Students share a plate:
  • one uses top row
  • one uses bottom row
• write a program on PS2

Student protocol:

• Make a 2-fold dilution series of the BSA
  • 0.5 mL water in each tube (except the first)
  • 1 mL BSA into first tube
  • .5 mL to next tube, mix, transfer to next, etc.
• Put 160 uL BSA into each well
• put 160 uL water in one well. (Why?)
• Add 40 uL Bradford
• Read plate

Data management

• Stupid Excel Tricks
• Art of the standard curve
• Make your own std curve.
• How would you estimate the concentration of protein in a solution that had an Abs of:
  • 0.5
  • 0.4
  • 3