User:Benjamin Friedel/Notebook/CHEM 471/2015/09/30: Difference between revisions
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span> | ||
|style="background-color: #F2F2F2" align="center"| | |style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} | ||
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Latest revision as of 01:14, 27 September 2017
Project name | Main project page Previous entry Next entry |
ObjectiveToday's objective was to complete the fluorescence analysis from yesterday (09/29/2015) as the blank standards were not run yet. Additionally, the objective was to analyze a-chymotrypsin degradation of lysozyme AuNP fibers over time at increased temperature (37˚C) through UV-Vis Bradford Analysis. ProtocolLysozyme Protease Degradation Fluorescence Analysis On 09/29/15 all of the lysozyme standards were analyzed, today the blank standard were analyzed using the same procedure. Bradford Analysis of Protease Degradation
Calculating Protease Volume in Reaction Mix
Calculating Buffer Volume in Reaction Mix
Blank Preparation
Bradford Analysis
DataFigure 1: Lysozyme Degradation by alpha-Chymotrypsin Absorbance by Wavelength Figure 1 below shows the Uv-Vis absorbance spectra for each of the timed Bradford samples corrected for blank, background from 700-800nm and isosbestic point at 535nm. To correct for the blank the blank spectra for each corresponding incubation time was subtracted from the lysozyme sample spectra for that same incubation time. To correct for background, the average absorbance between 700-800nm was calculated for each blank corrected spectra and subtracted from that spectra. To correct for isosbestic point absorbance at 535nm for each of the blank and background corrected spectra was subtracted from each respective spectra so that they all are equal to zero absorbance at 535nm. Figure 2: Lysozyme Degradation by 1uM alpha-Chymotrypsin Bradford Calibration Curve Figure 2 below shows the calibration curve of absorbance vs. protease incubation time in minutes and compares the samples to the blanks. For this data, the lysozyme sample spectra did not have their respective blank data subtracted from and were corrected for background and isosbestic point as shown above. Similarly, the blank data was corrected for background and isosbestic point as shown above. |