User:Matthew R Skorski/Notebook/471 - Exp BioChem/2016/03/30
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Objective
ProtocolMaking AuNP FibersWe made 10mL AuNP fiber samples in glass test tubes using the following reagents: We then added the following to each sample:
We put the samples in the oven and ran them on cycle P1. Measuring the UV-Vis Absorption of AuNP Fiber Supernatants After 2 Hour Incubation in RhodamineWe added the following amounts of Rhodamine to our lysozyme samples that we synthesized yesterday: We then covered our samples in Parafilm, inverted them gently to mix in the Rhodamine, and allowed them to incubate at room temperature in Rhodamine for 2 hours. After 2 hours, we spun down the samples, pipetted 3.5mL of the supernatants into polystyrene cuvettes, and measured the UV-Vis absorption of each. Measuring the UV-Vis Absorbance of Rhodamine as a Function of its Concentration in AuNP Fiber SupernatantSince we only used 9 samples for our 2 hour incubation, we used 10 of the remaining lysozyme samples that we synthesized yesterday in order to make a standard curve for the UV-Vis absorption of Rhodamine as a function of its concentration in the supernatant of our fiber samples. We pipetted the following volumes of Rhodamine from our 1390µM stock of Rhodamine into each of 10 samples in order to make standards: We then covered the samples in Parafilm, inverted them, and spun them down for 2 minutes. We pipetted 3.5mL of each supernatant into polystyrene cuvettes and measured the UV-Vis absorbance.
AnalysisAll of the BSA fibers we tried to synthesize yesterday failed. However, the lysozyme worked. We will use this higher ratio of gold:lysozyme in our samples moving forward. UV-Vis Absorption of AuNP Fiber Supernatants After 2 Hour Incubation in RhodamineCopied from Michael's notebook (as of April 6, 2016): Figure 2: The table above shows the raw and calculated data including the peak absorbance, the concentration that this absorbance corresponds to, the change in concentration after incubation, and the percent change in concentration.
Standard Curve for the UV-Vis Absorbance of Rhodamine as a Function of its Concentration in AuNP Fiber SupernatantAlso copied from Michael's notebook (as of April 6, 2016):
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