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

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## Add the correct amount of buffer such that the final volume is 1mL
## Add the correct amount of buffer such that the final volume is 1mL
# Take a UV-Vis (no less than 1 hour after they were produced).
# Take a UV-Vis (no less than 1 hour after they were produced).
## Use the plastic cuvettes.
# Make 2 blanks as well (800uL buffer and 200uL Assay reagent) and take it's UV spectrum. (400nm-800nm)
# Make 2 blanks as well (800uL buffer and 200uL Assay reagent) and take it's UV spectrum. (400nm-800nm)
# After you have finished one set, repeat the process (make new samples and new measurements)
# After you have finished one set, repeat the process (make new samples and new measurements)
# Make a calibration curve.
# Determine if you need to redo any data or sample prep.


We are also going to make Atomic Absorption standards for tomorrow!
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)
Using the gold AA/ICPMS standard solution make 5 new solutions (Note: Use water - NOT BUFFER - to make these solutions)

Revision as of 11:17, 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

Making buffer: 1L 50mM Tris 50mM NaCl pH 7.5

Should mass out:

  1. 6.057g Tris
  2. 2.922g NaCl

Actually measured:

  1. 6.0596g Tris
  2. 2.9525g NaCl

Gold solution for group: HAT to make nano particles

  1. Gold solution (HAuCl4·3H2O) 0.0050g in 0.0500mL water → 0.25mM



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