User:Puja Mody/Notebook/Chem 571: Gold Nanoparticles/2012/09/25: 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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#Sonicate the cells using sonication instrumentation. place the sonication device inside each of vials containing the cells for 30 seconds followed placing the vial in an ice bath for 30 seconds. repeat this sequence three times. This will shock and open up the cells. | #Sonicate the cells using sonication instrumentation. place the sonication device inside each of vials containing the cells for 30 seconds followed placing the vial in an ice bath for 30 seconds. repeat this sequence three times. This will shock and open up the cells. | ||
#centrifuge the cells at 18000 rpm and 4°C for 2 hours. This will separate the organelles of the cells from the proteins we want. | #centrifuge the cells at 18000 rpm and 4°C for 2 hours. This will separate the organelles of the cells from the proteins we want. | ||
#Meanwhile, filter the binding buffer and elution buffer in order to purify them using a vacuum system of filtration and filter paper with a membrane filter in which the holes measure 450nm. | #Meanwhile, filter the [[User:Puja Mody/Notebook/Chem 571: Gold Nanoparticles/2012/09/19| binding buffer]] and [[User:Puja Mody/Notebook/Chem 571: Gold Nanoparticles/2012/09/19| elution buffer]] in order to purify them using a vacuum system of filtration and filter paper with a membrane filter in which the holes measure 450nm. | ||
#Once the centrifuging is completed. Use the same vacuum filter system to separate the proteins from everything else in the vials (organelles, buffer, etc.) | #Once the centrifuging is completed. Use the same vacuum filter system to separate the proteins from everything else in the vials (organelles, buffer, etc.) | ||
# The protein was collected in falcon vials and refrigerated. | # The protein was collected in falcon vials and refrigerated. | ||
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*UV-Vis | *UV-Vis | ||
# New concentrations of the Tris buffer were added to the Gold/BSA solution. These solutions did not express any fibers, it was just purple solution. The UV-Vis was run twice on the samples with an hour in between each run. | # New concentrations of the Tris buffer were added to the Gold/BSA solution. These solutions did not express any fibers, it was just purple solution. The UV-Vis was run twice on the samples with an hour in between each run. | ||
==DATA/Calculations== | ==DATA/Calculations== |
Latest revision as of 22:04, 26 September 2017
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Separate ProteinsGoals
Procedure
DATA/Calculations
to make a 25mL stock solution of the tris buffer at a pH of 10 1mol/L x .025= .025 mol .025 mol x 121.14 g/mol = 3.0285 g of tris
link for excel File:TrisvaryingmolarUVVis.xlsx
Conclusions
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