User:Matthew R Skorski/Notebook/471 - Exp BioChem/2015/11/03: 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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###Protinase K concentration: (0.00115g)*(1mol/28,900g)*(1/0.001L)= 0.0000397 M Protinase K | ###Protinase K concentration: (0.00115g)*(1mol/28,900g)*(1/0.001L)= 0.0000397 M Protinase K | ||
####Proteinase K dilution: (39792 nM)*(0.05 mL Proteinase K) = (x nM)*(10 mL total dilution) => x = 199 nM | ####Proteinase K dilution: (39792 nM)*(0.05 mL Proteinase K) = (x nM)*(10 mL total dilution) => x = 199 nM | ||
####Proteinase K dilution: 0.05 mL of proteinase k solution was mixed with 9. | ####Proteinase K dilution: 0.05 mL of proteinase k solution was mixed with 9.95 mL of buffer | ||
###Amount of Proteinase K solution needed for 1mL with 1 nM concentration: M1*V1 = M2*V2 => (199 nM)*(V1) = (1 nM)*(1 mL) => V1 = 0.005 mL | ###Amount of Proteinase K solution needed for 1mL with 1 nM concentration: M1*V1 = M2*V2 => (199 nM)*(V1) = (1 nM)*(1 mL) => V1 = 0.005 mL | ||
###Amount of Buffer solution need to get to 1mL: (1mL total)-(0.005 mL Protinase K solution) = 0.995 mL buffer | ###Amount of Buffer solution need to get to 1mL: (1mL total)-(0.005 mL Protinase K solution) = 0.995 mL buffer | ||
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The graph below shows the peak for the absorbance at 600 nm. From 0 minutest to 75 minutes the aborbance rises, indicating more protein is free in solution. Then it drops at 135 minutes before rising again at 26 hour. More data points should be taken to ensure that the exact shape of the curve. | The graph below shows the peak for the absorbance at 600 nm. From 0 minutest to 75 minutes the aborbance rises, indicating more protein is free in solution. Then it drops at 135 minutes before rising again at 26 hour. More data points should be taken to ensure that the exact shape of the curve. | ||
[[Image:AMS_Bradford_1nM_Abs_600nm_vs_time.png|500px]] | [[Image:AMS_Bradford_1nM_Abs_600nm_vs_time.png|500px]] | ||
Latest revision as of 01:21, 27 September 2017
Project name | Main project page Previous entry Next entry |
ObjectiveThe objective of today was to run the Bradford Analysis of Protease Degradations for 1 nM Protinase K. DescriptionThe protocol is identical to the one used on 9/29/15 except the fibers were spun down at 300 rpm for 10 minutes, not 1500 rpm for 1 minute.
ResultsThe graph below shows the absorbance of the samples as function of the wavelength of incident light when corrected. The six samples were first corrected for by subtracting the absorbance of the superblank of buffer and Bradford reagents. Then the samples were corrected for again by subtracting the results of the first correction by the blanks for their corresponding time periods. Then, the isosbestic point at 535 nm from each samples was subtracted from all the wavelengths of that sample. The graph below shows the peak for the absorbance at 600 nm. From 0 minutest to 75 minutes the aborbance rises, indicating more protein is free in solution. Then it drops at 135 minutes before rising again at 26 hour. More data points should be taken to ensure that the exact shape of the curve. |