User:Mary Mendoza/Notebook/CHEM 571 Experimental Biological Chemistry I/2012/09/26: Difference between revisions
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* From the readings, the detection indicates that the absorbance is at 280; indicative of the presence of Histidine. However, the absorbance at 280 is not exclusive to Histidine alone but also corresponds to Tryptophan, Tyrosine, and Phenylalanine. | * From the readings, the detection indicates that the absorbance is at 280; indicative of the presence of Histidine. However, the absorbance at 280 is not exclusive to Histidine alone but also corresponds to Tryptophan, Tyrosine, and Phenylalanine. | ||
* The conductivity reading is sensitive to the amount of salt in the buffer. | * The conductivity reading is sensitive to the amount of salt in the buffer. | ||
* On the dialog box, pump > gradient > B > 100%. | * On the dialog box, pump > gradient > B > 100%. | ||
* Once the peak levels back to baseline, pump > gradient > B > 0%. This washes the system with A. | |||
* At Flowpath > injection valve > position > inject. Switch to position 1. This bypasses the column with the superloop's binding buffer flow at at 10 mL/min on inject. Then press end. | |||
* Sample 1 was loaded onto the super loop with the syringe at inject. The position was switched at position 3. The flow rate was at 5 mL/min with the gradient of B at 0%. From inject, the setting was switch to load. | |||
* The desired baseline should reach an absorbance of 280 and 250. Then hit pause. | |||
* The fraction is under the Frac 900 function in the dialog box. 5 mL fraction size was chosen. Then hit continue. | |||
* On the pump function, the load was set at 0% B after 25 mL the flow rate of 0 mL/min. was changed to 5 mL/min. | |||
* The same steps were done for sample 2. | |||
* The filtrate for sample 1 was collected from tubes 1-6 and sample 2 was collected from tubes 7-11. | |||
* Observing the graph below, the peak at fraction 2 indicates that most of the protein (ADA) was collected in this run. For sample 2, the protein filtrate was mostly collected at fraction 8. | |||
[[Image:ADA HisTrapE.PNG]] | |||
==Preparation of Au/BSA solution== | |||
* The nanoparticles were synthesized from 15 μM BSA with 2 mL of .0336 M Au diluted down to 10 mL of water. | |||
* The solutions were prepared with the same standard mole ratios as before. | |||
* Then, the solutions were placed to the thermo scientific for 4 h. at 80°C. | |||
Revision as of 06:49, 7 December 2012
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Purification of Proteins
Preparation of Au/BSA solution
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