User:Daniel-Mario Larco/Notebook/AU Biodesign Lab - 09/03/2013/2014/04/15: Difference between revisions
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== | ==Objective== | ||
* | * Test HRP pellet | ||
* Spin myoglobin and HB to see if it breaks apart | |||
* UV/Vis myoglobin and Hb, spin, test again | |||
==Procedure== | |||
#Make 0.1g/mol solutions of all three proteins in 50mM Tris buffer | |||
#Spin solutions at 19000 rpm for 3 hours | |||
#Test for HRP activity in the pellets | |||
##Make OPD solution (1mg/mL) | |||
##Make H2O2 solution | |||
###113uL 30% H2O2 in 887uL H2O | |||
## Make mixture to be tested | |||
### 2.025mL OPD solution | |||
### 0.75mL H2O2 solution | |||
### Take UV-Vis | |||
### Add 0.225mL | |||
==Data== | |||
[[Image:Screen Shot 2014-04-16 at 5.46.16 PM.png]] | |||
UV-Vis of Hemoglobin solution before Spin | |||
[[Image:Screen Shot 2014-04-16 at 5.46.24 PM.png]] | |||
UV-Vis of Myoglobin solution before Spin | |||
[[Image:Screen Shot 2014-04-16 at 5.47.09 PM.png]] | |||
UV-Vis of AuNP solution before spin | |||
*Used for concentration calculations and size | |||
[[Image:Screen Shot 2014-04-16 at 5.47.02 PM.png]] | |||
[[Image:Screen Shot 2014-04-16 at 5.46.34 PM.png]] | |||
[[Image:Screen Shot 2014-04-16 at 5.46.45 PM.png]] | |||
[[Image:Screen Shot 2014-04-16 at 5.46.53 PM.png]] | |||
The last three images represent the activity of the pellets of HRP:AuNP conjugates for the three methods | |||
==Notes== | |||
When the tubes were removed from the centrifuge, none of the solutions had separated, pelleted out. | |||
There was a homogenous mixture for all three. The tubes were stored in the fridge over night to test for activity. | |||
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Revision as of 02:19, 25 April 2014
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Objective
Procedure
DataUV-Vis of Hemoglobin solution before Spin UV-Vis of Myoglobin solution before Spin UV-Vis of AuNP solution before spin
The last three images represent the activity of the pellets of HRP:AuNP conjugates for the three methods NotesWhen the tubes were removed from the centrifuge, none of the solutions had separated, pelleted out. There was a homogenous mixture for all three. The tubes were stored in the fridge over night to test for activity. |