Bio 499CB (Loomis Cancer)

Submitted by kdorfman on Thu, 09/08/2022 - 20:59

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