User:Jamie Nunziata/Notebook/Protease Research/2015/11/04: Difference between revisions
(Autocreate 2015/11/04 Entry for User:Jamie_Nunziata/Notebook/Protease_Research) |
(fix raw html notebook nav) |
||
| (One intermediate revision by one other user not shown) | |||
| Line 2: | Line 2: | ||
|- | |- | ||
|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}}}}]]}} | ||
|- | |- | ||
| colspan="2"| | | colspan="2"| | ||
<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### --> | <!-- ##### DO NOT edit above this line unless you know what you are doing. ##### --> | ||
== | ==Objective== | ||
* | The purpose of this lab is to use Bradford Assay to measure AuNP fiber degradation from a solution of 1nM a-chymotrypsin protease | ||
==Procedure== | |||
7 samples of AuNP fibers were spun down at 300rpm for 10 minutes.These fibers will be used for our 23 hours, 120min, 90min, 60min, and 30min samples. A 1:200 dilution of our stock protease was made in Tris buffer. We pipetted 5µL of our 43.539µM stock solution of a-chymotrypsin into 995µL of Tris buffer. This brought the final concentration of the a-chymotrypsin to 0.20506µM. 4.88µL of the diluted a-chymotrypsin (in Tris) and 995.12µL of Tris buffer was added to each sample tube and a blank Eppindorf tube (no fibers), making the final solution 1nM a-chymotrypsin. | |||
To each cuvette, the following was added: | |||
*1650µL of Tris buffer | |||
*750µL of incubated 1µM sample (or blank) | |||
*600µL of diluted Bradford Assay in Tris buffer (1:3) | |||
A full lab procedure can be found in Nicole Bonan's notebook entry for [[User:Nicole_Bonan/Notebook/Chem_571_Lab_Notebook/2015/11/04|today]] | |||
Absorbance for each cuvette was measure via UV-Vis between the emission sprectrum of 400-800nm, and the data is recorded below | |||
==Data== | |||
[[Image:JMN_11_4_2015_CorrectedAborbanceSamples_Bradford.jpg]] | |||
[[Image:JMN_11_4_2015_BlankSample600nm_Bradford.jpg]] | |||
[[Image:JMN_11_4_2015_Sample600nm_Bradford.jpg]] | |||
. | |||
Latest revision as of 01:21, 27 September 2017
 Project name
|
 Main project page Previous entry Next entry
|
ObjectiveThe purpose of this lab is to use Bradford Assay to measure AuNP fiber degradation from a solution of 1nM a-chymotrypsin protease Procedure7 samples of AuNP fibers were spun down at 300rpm for 10 minutes.These fibers will be used for our 23 hours, 120min, 90min, 60min, and 30min samples. A 1:200 dilution of our stock protease was made in Tris buffer. We pipetted 5µL of our 43.539µM stock solution of a-chymotrypsin into 995µL of Tris buffer. This brought the final concentration of the a-chymotrypsin to 0.20506µM. 4.88µL of the diluted a-chymotrypsin (in Tris) and 995.12µL of Tris buffer was added to each sample tube and a blank Eppindorf tube (no fibers), making the final solution 1nM a-chymotrypsin.
A full lab procedure can be found in Nicole Bonan's notebook entry for today Absorbance for each cuvette was measure via UV-Vis between the emission sprectrum of 400-800nm, and the data is recorded below Data
| |
