User:Alicia Rasines Mazo/Notebook/CHEM-571 Experimental Biological Chemistry/2014/09/23: 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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*Run for 30 minutes at 200V | *Run for 30 minutes at 200V | ||
=====Develop/Stain your gel===== | =====Develop/Stain your gel===== | ||
# Place gel in Fixative Solution (40% methanol, 10% acetic acid, 50% water) for 30 minutes | # Place gel in Fixative Solution (40% methanol, 10% acetic acid, 50% water) for 30 minutes. | ||
#*Note: our gel was placed on top of Paul group's gel | |||
# Place gel in Stain Solution (0.025% (w/v) Coomassie Blue, 10% acetic acid, 90% water) for 1 hour | # Place gel in Stain Solution (0.025% (w/v) Coomassie Blue, 10% acetic acid, 90% water) for 1 hour | ||
#*Note: our gel was placed on the bottom of the container | |||
# Place gel in Destain Solution (10% acetic acid, 90% water) for 15 minutes | # Place gel in Destain Solution (10% acetic acid, 90% water) for 15 minutes | ||
#* Repeat this step with fresh destain solution 2 more times | #* Repeat this step with fresh destain solution 2 more times | ||
===Bradford analysis for protein content=== | ===Bradford analysis for protein content=== | ||
*Both for dialyzed and non-dyalized BSA colloid solutions | *Both for dialyzed and non-dyalized BSA colloid solutions | ||
*Also for outside solutions | |||
**25000 g/mol MW cut-off glycine soak | |||
**3500 g/mol MW cut-off glycine with colloid | |||
**3500 g/mol MW cut-off H<sub>2</sub>O soak | |||
**3500 g/mol MW cut-off NaCl soak | |||
**3500 g/mol MW cut-off glycine soak | |||
Note: protein seemed to hsve precipitated out of solution <br.> | |||
Bradford analysis of outside solutions: | |||
#600μL of outside solution | |||
#200μL Bradford reagent | |||
# 200μL Tris/NaCl buffer | |||
Run UV-Vis from 400 to 800 nm. |
Latest revision as of 00:19, 27 September 2017
Project name | Main project page Previous entry Next entry |
Task for September 2014
Procedure followed as described by Dr. Hartings on Sept. 25 2013 ElectrophoresisPrepare the Gel and Assemble the Electrophoresis Cell
Note: 200mL of running buffer (10x SDS-PAGE running buffer 30.3 g Tris, 144.1g Clycine, 10g SDS, water to 1L) were diluted to 2 L. This amount was necessary in order to fill up the inner and outer buffer chambers Prepare and Load Samples
Perform Electrophoresis
Develop/Stain your gel
Bradford analysis for protein content
Note: protein seemed to hsve precipitated out of solution <br.> Bradford analysis of outside solutions:
Run UV-Vis from 400 to 800 nm. |