Difference between revisions of "UV Vis"
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==UV-Visible Spectoscopy== | ==UV-Visible Spectoscopy== | ||
| − | UV-Vis is used to measure absorption spectroscopy in the ultraviolet-visible spectral region. We are using the UV-Vis to measure the activity of peroxidases, starting with HRP to develop a standard protocol | + | UV-Vis is used to measure absorption spectroscopy in the ultraviolet-visible spectral region. We are using the UV-Vis to measure the activity of peroxidases, starting with HRP to develop a standard protocol. We used the kinetics mode of the UV-Vis to gather our data. |
'''Steps to Run a Peroxidase through the UV-Vis | '''Steps to Run a Peroxidase through the UV-Vis | ||
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1) Turn on UV-Vis | 1) Turn on UV-Vis | ||
2) Open the UV-Vis software on the computer monitor | 2) Open the UV-Vis software on the computer monitor | ||
| − | 3)Turn on Kinetics Mode | + | 3) Turn on Kinetics Mode |
a) Set wavelength to the desired absorption (ours was 436 nm for Guaiacol) | a) Set wavelength to the desired absorption (ours was 436 nm for Guaiacol) | ||
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4) Select time-based measurements and create a name for each set of data before running the sample | 4) Select time-based measurements and create a name for each set of data before running the sample | ||
| − | 5)Run a blank with DI water before running a sample | + | 5) Run a blank with DI water before running a sample |
6) Fill a cuvette with the assay and place the cuvette in the UV-Vis | 6) Fill a cuvette with the assay and place the cuvette in the UV-Vis | ||
Latest revision as of 23:07, 30 June 2020
UV-Visible Spectoscopy
UV-Vis is used to measure absorption spectroscopy in the ultraviolet-visible spectral region. We are using the UV-Vis to measure the activity of peroxidases, starting with HRP to develop a standard protocol. We used the kinetics mode of the UV-Vis to gather our data.
Steps to Run a Peroxidase through the UV-Vis
1) Turn on UV-Vis
2) Open the UV-Vis software on the computer monitor
3) Turn on Kinetics Mode
a) Set wavelength to the desired absorption (ours was 436 nm for Guaiacol)
b) Set a cycle time - the lower the cycle time the more data collected (we used 0.5 seconds)
c) Set a run time - HRP does not show significant data past a minute (so we used 60 seconds)
4) Select time-based measurements and create a name for each set of data before running the sample
5) Run a blank with DI water before running a sample
6) Fill a cuvette with the assay and place the cuvette in the UV-Vis
7)Add desired amount of peroxidase to the assay and mix with a plastic transfer pipette
8)Select the sample button on the monitor