User:Alexander Cvitan/Notebook/Experimental Biological Chemistry Lab/2014/01/29: Difference between revisions

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===pH===
===pH===
====Creating 1 M Buffer Solutions====
====How we made each 1 M Buffer Solution====
*Please note that all of the following dilutions were calculated via an online buffer calculator found [[http://www.biomol.net/en/tools/buffercalculator.htm| HERE.]]
*Please note that all of the following dilutions were calculated via an online buffer calculator found [[http://www.biomol.net/en/tools/buffercalculator.htm| HERE.]]
=====Citric Acid: pH=2.99=====
=====Citric Acid: pH=2.99=====

Revision as of 11:50, 1 February 2014

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Purpose

  • Make stock buffer solutions at specific pH's.
  • See the effect of the addition of these buffer solutions on Lysozyme AuNP fiber formation.

Procedure

  • Mix all test tubes in order to re-suspend the fibers in the solution. (Not to dissolve but to evenly distribute the fibers in the solution.)
  • Take 1 mL of solution from each test tube and place it in a separate 1.5 mL tube. (These aliquots will initially be used in order test each variable.)

Chelators

Creating Stock Solutions of Chelators

File:1.29.data.chelator.stock

Adding Chelators to Lysozyme-AuNP Solutions

  • Add 0.5 mL of each chelator stock solution to each 1.5 mL aliquot.
  • Record effects of each chelator on each solution.

pH

How we made each 1 M Buffer Solution

  • Please note that all of the following dilutions were calculated via an online buffer calculator found [HERE.]
Citric Acid: pH=2.99
  • Add 42.02 g of citric acid in 180 mL distilled water.
  • Titrate to pH 3.0 with monovalent strong acid/base if necessary.
  • Fill up to 200 mL with distilled water.
Phosphate: pH=6.81
  • Add 0.0908 mol of sodium phosphate monobasic to 0.1091 mol of sodium phosphate dibasic.
  • Fill up to 200 mL with distilled water.
Tris
  • Dissolve 24.228 g Tris base in 180 mL distilled water.
  • Titrate to pH 9.0 with monovalent strong acid/base if necessary.
  • Fill up to 200 mL with distilled water.
MES: pH=5.20
  • Dissolve 39.04 g MES in 180 mL distilled water.
  • Titrate to pH 5.2 with monovalent strong acid/base if necessary.
  • Fill up to 200 mL with distilled water.
HEPES: pH=7.05
  • Dissolve 47.66 g HEPES in 180 mL distilled water.
  • Titrate to pH 7 with monovalent strong acid/base if necessary.
  • Fill up to 200 mL with distilled water.
CHES: pH=8.97
  • Dissolve 41.46 g CHES in 180 mL distilled water.
  • Titrate to pH 9 with monovalent strong acid/base if necessary.
  • Fill up to 200 mL with distilled water.

Adding Buffers to Lysozyme-AuNP Solutions

  • Dilute the 1 M solutions to 100 mM, 10 mM, and 1 mM.
  • Add 1 mL of each to the lysozyme-AuNP solutions.
  • Let sit overnight and record results.



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