User:Beatriz Gimenez De C./Notebook/571/2014/09/10: Difference between revisions
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*vortex tubes and let them sit for 5 min. | *vortex tubes and let them sit for 5 min. | ||
*Obtain a UV-VIS spectrum (between 400 and 800 nm). | *Obtain a UV-VIS spectrum (between 400 and 800 nm). | ||
*Make 2 duplicate blanks (1 mL Tris/NaCl | *Make 2 duplicate blanks (1 mL Tris/NaCl buffer and 200 μL BRADFORD reagent + 800 μL buffer) | ||
*repeat the process using Lysozyme. | *repeat the process using Lysozyme. | ||
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==Calculations== | ==Calculations== | ||
* used absorbance values to calculate the concentrations | |||
* personal note : table yet to be uplaoded ( need to ask how it is done) | |||
==Note== | |||
* Worked with Paul because Monika was absent. | |||
Revision as of 09:59, 18 September 2014
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Task
- Prepare 50 mL of a standard saline solution (0.9 wt-% NaCl). Store in a 45 mL Falcon tube.
- Prepare 50 mL of a 50 mM Tris (not Tris-HCl) 50 mM NaCl solution. Store in a 45 mL Falcon tube.
- Prepare a stock solution of BSA that is roughly 5 mg in 5 mL of saline.
- Record UV-VIS spectra between 200 nm and 800 nm.
- 6 - 8 standard solutions 1 μg/mL and 20 μg/mL. Using determined volume of stock solution to add to 1.5 mL centrifuge tube. Add 200 μL of the Bio-Rad Protein Assay reagent. Use 1:4 concentrate diluted with water. Add to the solution Tris/NaCl until 1 mL volume is reached.
- vortex tubes and let them sit for 5 min.
- Obtain a UV-VIS spectrum (between 400 and 800 nm).
- Make 2 duplicate blanks (1 mL Tris/NaCl buffer and 200 μL BRADFORD reagent + 800 μL buffer)
- repeat the process using Lysozyme.
Results
- UV-Vis data from the 6 serial dilutions of BSA (1 μg/mL, 2 μg/mL, 5 μg/mL, 10 μg/mL, 15 μg/mL, 20 μg/mL)
- personal note : Graph and table yet to be uplaoded ( need to ask how it is done)
- UV-Vis data from the 6 serial dilutions of Lysozyme (1 μg/mL, 2 μg/mL, 5 μg/mL, 10 μg/mL, 15 μg/mL, 20 μg/mL)
- personal note : Graph and table yet to be uplaoded ( need to ask how it is done)
Calculations
- used absorbance values to calculate the concentrations
- personal note : table yet to be uplaoded ( need to ask how it is done)
Note
- Worked with Paul because Monika was absent.
