User:Benjamin Friedel/Notebook/CHEM 471/2015/10/28: 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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'''Incubation Mix Preparation''' | '''Incubation Mix Preparation''' | ||
# To the Ocean Optics Cuvette, the 1mL total resuspended fiber samples was added as well as the volume of phosphate buffer as shown above. | # To the Ocean Optics Cuvette, the 1mL total resuspended fiber samples was added as well as the volume of phosphate buffer as shown above. | ||
## Experimental and save parameters were set to leave | ## Experimental and save parameters were set to leave 30sec between scans and to only scan for 24 hours. However, after 2 hours of collecting data under these parameters, the parameters were changed to 20min between scans from that time to the end of the initial 24 hour time period. Additionally, the scan ranged from 190.96nm-891.81nm. | ||
## To begin the experiment, the lower save file button and then the play button were clicked, creating an immediate first save file. | ## To begin the experiment, the lower save file button and then the play button were clicked, creating an immediate first save file. | ||
# Then, immediately after the second save file was generated after 2min, the volume of protease (alpha chymotrypsin) as shown above was added to the sample. | # Then, immediately after the second save file was generated after 2min, the volume of protease (alpha chymotrypsin) as shown above was added to the sample. |
Latest revision as of 01:20, 27 September 2017
Project name | Main project page Previous entry Next entry |
ObjectiveOcean Optics Spectrophotometry was used to measure absorbance of an AuNP fibers sample over time as it was incubated with the protease, alpha-chymotrypsin. This was performed to track how alpha-chymotrypsin digests lysozyme based AuNP fibers. ProtocolAuNP Fiber Sample Preparation
Protease Preparation
Incubation Mix Preparation
DataData Processing In this experiment, the blank or control data set is considered to be the first data point collected after the initial data point collection. To correct for this blank data (without protease), the absorbance values at each wavelength were subtracted from the absorbance data for each 2 minute measurement after addition of protease (alpha chymotrypsin). Then, to correct for instrumental noise, the absorbance values for the last 60nm for each 2min scan were averaged individually and subtracted from that individual scan.
Figure 1 above shows the absorbance values for each scan after the first 2min at 530nm are shown as time progressed. This is because AuNP fibers were previously determined to have an absorbance max at 530nm, so their breakdown into solution upon digestion of AuNP fibers by alpha-chymotrypsin was monitored. This figure, in contrast to data collected on 10/07/15 shows that before the protease was added, lower absorbance. After protease addition absorbance increases and plateau's around A=.11. |