User:Javier Vinals Camallonga/Notebook/Javier Vinals notebook/2014/01/29

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Objective

Today we are going to create stock Solutions of different pH’s, in order to see how pH affects the formation of fibers.

Procedure

In order to create the buffer Solutions, we used a buffer calculador, where we were able to find the buffers we wanted and the amounts of each solvent to be added onto water. We made 1 M Solutions, and then we will dilute them to 100 mM and to 10mM, and see the effect on the solution depending on the concentration of the solution.

Creating 1 M Buffer Solutions

Citric Acid: pH=2.99

  1. Add 42.02 g of citric acid in 180 mL distilled water.
  2. Titrate to pH 3.0 with monovalent strong acid/base if necessary.
  3. Fill up to 200 mL with distilled water.

Phosphate: pH=6.81

  1. Add 0.0908 mol of sodium phosphate monobasic to 0.1091 mol of sodium phosphate dibasic.
  2. Fill up to 200 mL with distilled water.

Tris

  1. Dissolve 24.228 g Tris base in 180 mL distilled water.
  2. Titrate to pH 9.0 with monovalent strong acid/base if necessary.
  3. Fill up to 200 mL with distilled water.

MES: pH=5.20

  1. Dissolve 39.04 g MES in 180 mL distilled water.
  2. Titrate to pH 5.2 with monovalent strong acid/base if necessary.
  3. Fill up to 200 mL with distilled water.

HEPES: pH=7.05

  1. Dissolve 47.66 g HEPES in 180 mL distilled water.
  2. Titrate to pH 7 with monovalent strong acid/base if necessary.
  3. Fill up to 200 mL with distilled water.

CHES: pH=8.97

  1. Dissolve 41.46 g CHES in 180 mL distilled water.
  2. Titrate to pH 9 with monovalent strong acid/base if necessary.
  3. Fill up to 200 mL with distilled water.

Adding Buffers to Lysozyme-AuNP Solutions

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