User:Eleni N. Kalivas/Notebook/CHEM-571/2013/10/22

Project name

Main project page Previous entry      Next entry

Objective The objective of today's lab is to synthesize AuNPs for reactivity tests tomorrow. Also to determine the Au;Protein ratio that is required for fiber formation of HRP and for pepsin. Protocol The following has been taken from Matt Harting's lab notebook. Synthesis of HRP-AuNPs and Pepsin-AuNPs Into test tube that will contain a total of 5mL add Au solution such that the final concentration is 0.25mM Protein solution such that the final concentration reflects the following Au:Protein ratios 30 50 70 90 110 130 150 170 190 210 Water up to 5mL HRP-AuNP and Pepsin-AuNP 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 Data Absorbance of BSA-AuNPs was determined for future comparison of the activity of protein-AuNPs. Absorbance spectra of a 1/10 dilution of BSA-AuNPs Absorbance of all starting materials used for synthesis of AuNPs today for future comparison Absorbance spectra of a 1/10 dilution of 2.52mM gold in water Absorbance spectra of 1/2 dilution of 15.3μM HRP Absorbance spectrum of 1/10 dilution of 15.6μM pepsin No absorbance peak was observed for pepsin. This was expected due to the lack of color of the pepsin stock solution.