User:Helen L. Slucher/Notebook/CHEM 571/2013/09/10: Difference between revisions

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# 5 ug/mL
# 5 ug/mL


==Data==
==Data (Trial 1)==
Making buffer:
*[[Image:Screen shot 2013-10-12 at 2.49.52 PM.png]]
1L 50mM Tris 50mM NaCl pH 7.5
*[[Image:Screen shot 2013-10-12 at 2.50.56 PM.png]]


Should mass out:
==Data (Trial 2)==
# 6.057g Tris
*[[Image:Screen shot 2013-10-12 at 2.50.19 PM.png]]
# 2.922g NaCl
*[[Image:Screen shot 2013-10-12 at 2.51.18 PM.png]]
 
Actually measured:
# 6.0596g Tris
# 2.9525g NaCl
 
Gold solution for group: HAT to make nano particles
 
# Gold solution (HAuCl<sub>4</sub>·3H<sub>2</sub>O) 0.0050g in 0.0500mL water → 0.25mM




Revision as of 11:52, 12 October 2013

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Objective

Make calibration curves using the Bradford Assay.

Procedure

  1. Make a stock solution that is roughly 10mg protein in 2mL of buffer.
  2. Calculate your actual concentration
  3. Make 6 standard solutions (1mL each) between 1μg/mL and 10μg/mL
    1. Determine the appropriate volume of your stock to use (for the proper final concentration in 1mL) and add that volume to an eppendorf tube
    2. Add 200μL of the Bio-Rad Protein Assay reagent
    3. Add the correct amount of buffer such that the final volume is 1mL
  4. Take a UV-Vis (no less than 1 hour after they were produced).
  5. Make 2 blanks as well (800uL buffer and 200uL Assay reagent) and take it's UV spectrum. (400nm-800nm)
  6. After you have finished one set, repeat the process (make new samples and new measurements)

Make Atomic Absorption standards for tomorrow!

Using the gold AA/ICPMS standard solution make 5 new solutions (Note: Use water - NOT BUFFER - to make these solutions)

  1. 25 ug/mL
  2. 20 ug/mL
  3. 15 ug/mL
  4. 10 ug/mL
  5. 5 ug/mL

Data (Trial 1)

Data (Trial 2)



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