User:Elaine Marie Robbins/Notebook/CHEM-496/2011/10/25: Difference between revisions
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|style="background-color: #EEE"|[[Image:BDLlogo_notext_lr.png|128px]]<span style="font-size:22px;"> | |style="background-color: #EEE"|[[Image:BDLlogo_notext_lr.png|128px]]<span style="font-size:22px;"> SDS-PAGE</span> | ||
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html> </html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}} | |style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html> </html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}} | ||
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==Objective== | ==Objective== | ||
To run an SDS-PAGE to determine whether or not MBP-intein fusion proteins are being expressed cells comparable to the cells frozen on [[User:Elaine_Marie_Robbins/Notebook/CHEM-496/2011/09/28|09/28/11]]. | |||
==Description== | ==Description== | ||
# Make solutions for a 12% discontinuous gel using the [http://ergo.berkeley.edu/be115/protean3.pdf BioRad Mini-PROTEAN instructions]. | |||
# Set resolving gel solution in gel holder with methanol on top to make the top of the gel flat. | |||
# Allow gel to sit for 20 min. | |||
# Remove methanol. | |||
# Add stacking gel solution. | |||
# Load gels into SDS-PAGE. | |||
# Load ladder and protein solutions into gel. | |||
# Run SDS-PAGE. | |||
# Stain gels with Coomassie Brilliant Blue R250 solution. | |||
## Dissolve 0.2555 g Coomassie Brilliant Blue R250 in 90 mL of methanol:water (1:1 v/v) and 10 mL of glacial acetic acid. | |||
## Filter solution through a Whatman No. 1 filter. | |||
# Place on shaker overnight. | |||
==Data== | ==Data== | ||
Protein visibly traveled on gel after SDS-PAGE. | |||
==Notes== | ==Notes== | ||
Protein solutions used were from group 2: | |||
* [[User:Samiye_Yaman/Notebook/Lab_571|Samiye Yaman]] | |||
* [[User:Elizabeth_Ghias/Notebook/Experimental_Biological_Chemistry| Elizabeth Ghias]] | |||
Revision as of 09:29, 26 October 2011
SDS-PAGE | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> |
ObjectiveTo run an SDS-PAGE to determine whether or not MBP-intein fusion proteins are being expressed cells comparable to the cells frozen on 09/28/11. Description
DataProtein visibly traveled on gel after SDS-PAGE. NotesProtein solutions used were from group 2:
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