# Difference between revisions of "User:Zachary I. Mendel/Notebook/Zacks Notebook/2013/10/22"

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## Objective

1. Synthesize Pepsin-AuNPs for reactivity tests tomorrow
2. Determine the Au:Protein ratio required for fiber formation for pepsin and HRP

## Description

1. Synthesis of HRP-AuNPs and Pepsin-AuNPs
1. Into test tube that will contain a total of 5mL add
2. Au solution such that the final concentration is 0.25mM
3. Protein solution such that the final concentration reflects the following Au:Protein ratios
1. 30
2. 50
3. 70
4. 90
5. 110
6. 130
7. 150
8. 170
9. 190
10. 210
4. Water up to 5mL

## Data

Stock Solutions

1. HRP
1. 67.3mg in 100mL of water ---> 15.3 μM
2. Pepsin
1. 54.4mg in 100mL of water ---> 15.7 μM
3. Gold
1. Need 50mL of 2.5mM
2. 49.7mg of HAuCl4·3H2O in 50mL water --> 2.52 mM

## Notes

Table 1. Preparation of Protein-AuNP solutions

 Ratio Amount of gold added(mL) concentration HRP(M) concentration Pepsin(M) Volume HRP(mL) Volume Pepsin(mL) Volume Water HRP (mL) Volume Water Pepsin (mL) 30 0.496031746 8.33333E-06 8.33333E-06 2.723311547 2.653927813 1.780656707 1.850040441 50 0.496031746 0.000005 0.000005 1.633986928 1.592356688 2.869981326 2.911611566 70 0.496031746 3.57143E-06 3.57143E-06 1.16713352 1.137397634 3.336834734 3.36657062 90 0.496031746 2.77778E-06 2.77778E-06 0.907770516 0.884642604 3.596197738 3.61932565 110 0.496031746 2.27273E-06 2.27273E-06 0.742721331 0.723798494 3.761246923 3.780169759 130 0.496031746 1.92308E-06 1.92308E-06 0.628456511 0.61244488 3.875511743 3.891523374 150 0.496031746 1.66667E-06 1.66667E-06 0.544662309 0.530785563 3.959305945 3.973182691 170 0.496031746 1.47059E-06 1.47059E-06 0.480584391 0.468340202 4.023383863 4.035628052 190 0.496031746 1.31579E-06 1.31579E-06 0.42999656 0.419041234 4.073971694 4.08492702 210 0.496031746 1.19048E-06 1.19048E-06 0.389044507 0.379132545 4.114923747 4.124835709

• The varying ratios will be used to determine the ideal concentrations for the preparation of fibers of unraveled proteins.

• Absorbance of BSA-AuNPs was determined for future comparison of the activity of protein-AuNPs.

Figure 1. Absorbance spectra of a 1/10 dilution of BSA-AuNPs

• Absorbance of all starting materials used for synthesis of AuNPs today for future comparison

Figure 2. Absorbance spectra of a 1/10 dilution of 2.52mM gold in water

Figure 3. Absorbance spectra of 1/2 dilution of 15.3μM HRP

Figure 4. Absorbance spectrum of 1/10 dilution of 15.6μM pepsin

• No absorbance peak was observed for pepsin.