User:Floriane Briere/Notebook/CHEM-496/2012/02/14: Difference between revisions

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==Protocol==
==Protocol==


# Changing the pH of the solution
# Changing the pH of the solution (to 8.50)
##  
## Control solution:
### Initial solution: pH = 3.98 and volume = 10mL
### Add 321µL of buffer
### Final solution: pH = 8.50 and volume = 10.32mL
## Fibers solution (BSA/Gold = 166):
### Initial solution: pH = 3.17 and volume = 10mL
### Add 1.5mL of buffer
### Final solution: pH = 8.51 and volume = 11.5mL
## Purple solution (BSA/Gold = 70):
### Initial solution: pH = 3.50 and volume = 10mL
### Add 600µL of buffer
### Final solution: pH = 8.50 and volume = 10.6mL


* Preparation of the dye solution
# Preparation of the dye solution
## Tube 1 (for control solution): 2.0mg of 5(6)-Carboxynaphthofluorescein, N-succinimidyl ester (473.51 g/mol) + 160µL of DMSO
## Tube 2 (for 166 solution): 2.0mg of 5(6)-Carboxynaphthofluorescein, N-succinimidyl ester (473.51 g/mol) + 160µL of DMSO
## Tube 3 (for 70 solution): 2.4mg of 5(6)-Carboxynaphthofluorescein, N-succinimidyl ester (473.51 g/mol) + 160µL of DMSO


* Reaction with the Golds NPs solution (BSA/Gold = 70)
# Tagging the peptides with the dye:
* Reaction with the Golds NPs solution (BSA/Gold = 166)
## Tube 1 (control solution): 160µL of dye + 10.32mL of control solution
* Reaction with the control solution
## Tube 2 (166 solution): 160µL of dye + 11.5mL of fibers solution
## Tube 3 (70 solution): 160µL of dye + 10.6mL of purple solution
## Incubation for 24 hours at room temperature in the shaker


==Data==
==Data==

Revision as of 12:47, 14 February 2012

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Objective

Protocol

  1. Changing the pH of the solution (to 8.50)
    1. Control solution:
      1. Initial solution: pH = 3.98 and volume = 10mL
      2. Add 321µL of buffer
      3. Final solution: pH = 8.50 and volume = 10.32mL
    2. Fibers solution (BSA/Gold = 166):
      1. Initial solution: pH = 3.17 and volume = 10mL
      2. Add 1.5mL of buffer
      3. Final solution: pH = 8.51 and volume = 11.5mL
    3. Purple solution (BSA/Gold = 70):
      1. Initial solution: pH = 3.50 and volume = 10mL
      2. Add 600µL of buffer
      3. Final solution: pH = 8.50 and volume = 10.6mL
  1. Preparation of the dye solution
    1. Tube 1 (for control solution): 2.0mg of 5(6)-Carboxynaphthofluorescein, N-succinimidyl ester (473.51 g/mol) + 160µL of DMSO
    2. Tube 2 (for 166 solution): 2.0mg of 5(6)-Carboxynaphthofluorescein, N-succinimidyl ester (473.51 g/mol) + 160µL of DMSO
    3. Tube 3 (for 70 solution): 2.4mg of 5(6)-Carboxynaphthofluorescein, N-succinimidyl ester (473.51 g/mol) + 160µL of DMSO
  1. Tagging the peptides with the dye:
    1. Tube 1 (control solution): 160µL of dye + 10.32mL of control solution
    2. Tube 2 (166 solution): 160µL of dye + 11.5mL of fibers solution
    3. Tube 3 (70 solution): 160µL of dye + 10.6mL of purple solution
    4. Incubation for 24 hours at room temperature in the shaker

Data

  • Add data and results here...

Notes

This area is for any observations or conclusions that you would like to note.


Use categories like tags. Change the "Course" category to the one corresponding to your course. The "Miscellaneous" tag can be used for particular experiments, as instructed by your professor. Please be sure to change or delete this tag as required so that the categories remain well organized.

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