User:Khyra A. Neal/Notebook/Chem 571/2014/09/10: 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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**Purity= [UV Measured] / [Mass Measured] *100 | **Purity= [UV Measured] / [Mass Measured] *100 | ||
***1.736 X 10<sup>-5</sup>M / 1.5204 X 10<sup>-5</sup>M *100 = 114% | ***1.736 X 10<sup>-5</sup>M / 1.5204 X 10<sup>-5</sup>M *100 = 114% | ||
'''NOTE''' The extinction coefficient for BSA at 280 nm was found in literature to be 43,824. The percent purity of BSA was found to be greater than 1 possibly indicating that the polystyrene cuvettes weren't completely clean or BSA did not dissolve completely in solution and stuck to the cuvette. Because the Bradford reagent has peaks at 460 nm and 630 nm and the Bradford-protein complex has a peak near 600, there will be significant overlap. A difference spectra is constructed to make up for the difference and is shown below: | '''NOTE''' The extinction coefficient for BSA at 280 nm was found in literature to be 43,824. The percent purity of BSA was found to be greater than 1 possibly indicating that the polystyrene cuvettes weren't completely clean or BSA did not dissolve completely in solution and stuck to the cuvette. Because the Bradford reagent has peaks at 460 nm and 630 nm and the Bradford-protein complex has a peak near 600, there will be significant overlap. A difference spectra is constructed to make up for the difference and is shown below: | ||
[[Image:BSADifferenceSpectra.jpg]] | [[Image:BSADifferenceSpectra.jpg]] | ||
Latest revision as of 00:18, 27 September 2017
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September 10, 2014Bradford AssayPrepare Stock Solutions
UV-Vis AnalysisRecord UV-Vis Spectrum for stock solutions (BSA, Saline Solution, and Tris Buffer)
NOTE ALL UV-Vis spectra were run using polystyrene cuvettes. Solutions were discarded in waste bottle and polystyrene cuvettes were placed in a wash tube for cleaning. Data Analysis1. Purity of BSA stock solution Based on the spectra above, the absorbance of BSA at 280 nm = 0.761
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