Magnetite hydrogel synthesis

Magnetite- Protein Hydrogel Synthesis Protocol

1. Begin the heating of the water in the sonicator to 69 degrees Celsius. Do not turn on the sonication function at this time
2. Before beginning preparation of stock solutions be sure to wear appropriate safety gear (goggles, lab coat, gloves)

• Preparation of Stock Solutions

1. Weigh out the following compound and make stock solutions at the following volumes in volumetric flasks using deionized water. After the compound is dissolved in solution, transfer the stock solutions into falcon tubes. Use different volumetric flasks for preparation of hemoglobin and potassium nitrate.

Preparation of FeSO4 stock	'	'
Molecular Weight (g/mol)	278.01
mass(g)	2.085075
volume stock (L)	0.025	Concentration FeSO4 want in each solution (M)
Concentration (M)	0.3	0.03
Preparation of KOH stock
Molecular Weight (g/mol)	56.11
mass(g)	1.5
volume stock (L)	0.025	Concentration of KOH want in each solution (M)
Concentration (M)	1.069328106	0.1
Preparation of KNO3
Molecular Weight (g/mol)	101.1
mass(g)	9.099
volume stock (L)	0.1	Concentration of KNO3 want in each solution (M)
Concentration (M)	0.9	0.4
Preparation of hemoglobin Stock solution
Molecular weight (g/mol)	64500
mass hemglobin (g)	0.0995
volume solution (L)	0.1
Concentration (M)	1.54264E-05

Total volume of samples: 20mL

Ratio	Concentration Hemoglobin	Volume Hemoglobin Added (mL)	amount of FeSO4 added(mL)	amount of KOH added (mL)	amount of KNO3 added (mL)	volume Water added (mL)
16500	1.81818E-06	2.357240749	2	1.870333333	8.888888889	4.883537029

• Preparation of Samples

1. Prior to making the magnetite samples, label/ number 7 falcon tubes (1-7) with the date and ratio of sample being made. Weigh the mass of each of the falcon tubes and record in lab notebook.
2. Prepare 7 samples at a ratio of 16500:1 Magnetite- Hemoglobin. You want the total volume of each sample to be 20mL.
3. Pipette the necessary volumes of each stock solution into each of the 7 falcon tubes. Use deionized water to bring the total volume in the falcon tubes to 20mL.
   1. Be sure to add the Iron stock solution LAST.
   2. Store remaining stock solutions for later use
   3. Ensure that the hot water bath is at 69 degrees. Place the samples in the hot water bath (make sure that the samples are completely submerged) and turn on the sonication function. Set the timer for 90 minutes.
4. After 90 minutes, return and set the timer once again for another 90 minutes. Therefore the samples will have been sonicating for a total of 3 hours at 69 degrees Celsius.
5. While the samples are in the hot water bath, clean up your workstation and all necessary glassware. The hemoglobin volumetric flask may need to be placed in a base bath in order to remove particulate from the inside.