User:Moira M. Esson/Notebook/CHEM-571/2013/10/16: Difference between revisions
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*Catalytic activity was tested using UV/vis spectroscopy. | *Catalytic activity was tested using UV/vis spectroscopy. | ||
*3mL sample of luminol, H2O2 and buffer was placed in cuvette with micro-stir bar. After 1min., 100μL HRP-Au was added. | *3mL sample of luminol, H2O2 and buffer was placed in cuvette with micro-stir bar. After 1min., 100μL HRP-Au was added. | ||
*A spectrum was taken every | *A spectrum was taken every 15 seconds for ten minutes. | ||
<br> | <br> | ||
'''Sample preparation''' | '''Sample preparation''' | ||
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*No activity of the HRP was observed (see figure below). | *No activity of the HRP was observed (see figure below). | ||
<br> | <br> | ||
==UV/vis of Citrate and BSA samples== | ==UV/vis of Citrate and BSA samples== | ||
*A new BSA-Au sample was prepared because the protein had crashed out of solution. The BSA solution was prepared following the protocol described on [[User:Moira M. Esson/Notebook/CHEM-571/2013/08/28|08/28/2013]]. The absorbance of the BSA solution will be determined during the next meeting. | *A new BSA-Au sample was prepared because the protein had crashed out of solution. The BSA solution was prepared following the protocol described on [[User:Moira M. Esson/Notebook/CHEM-571/2013/08/28|08/28/2013]]. The absorbance of the BSA solution will be determined during the next meeting. |
Revision as of 14:41, 21 October 2013
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Objectives
Catalytic activity testing
Buffer:0.6140g Tris in 1L, pH set to 8 with HCl ---> 5.1mM Luminol:Dissolve 12.9mg luminol in 300uL of DMSO and add to 50mL buffer---> 1.46mM H2O2: 312uL 30% H2O2 into 100mL buffer ---> Should be 45mM
Calculations for preparation of sample: Luminol stock solution preparation 5mL 7.26E-5 M luminol: 0.261mL luminol stock added to 5mL buffer. H2O2 stock preparation of 5mL 0.0038M(this is 50x more concentrated): 0.169mL stock in 5mL buffer.
UV/vis of Citrate and BSA samples
Preparation of BSA stock solution 0.025g BSA in 25mL tris buffer(described above)-->15μM BSA
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