User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/09/12

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==Purpose==
==Purpose==
* To check if solutions made on [[User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/09/11|2012/09/11]] has produced fibers
* To check if solutions made on [[User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/09/11|2012/09/11]] has produced fibers
-
* To mix supernatant of sample with fiber pellet with different concentration of Tris buffer at pH 10 and check re-suspension
+
* To mix supernatant with pellet in each sample with the addition of different concentration of Tris buffer at pH 10 and check re-suspension
-
* Prepare the ratio 80 Au/BSA sample for peak shift testing next week
+
==Procedure==
==Procedure==
 +
* Sets of Au/BSA solutions with different ratios were remade from ratio 80 to ratio 170. For details with the volume, see [[User:Dhea Patel/Notebook/Experimental Biological Chemistry Notebook/2012/09/12|Dhea's Notebook]]. The sets of solutions were incubated in 85°C for four hours. After four hours of incubation, 1mL of supernatant were measured for absorption activity under UV-vis from 700nm to 200nm. See [[User:Dhea Patel/Notebook/Experimental Biological Chemistry Notebook/2012/09/12|Dhea's Notebook]] for details on absorption of each Au/BSA solutions after 4 hours of incubation.
 +
* 100mM Tris buffer at pH 10 were diluted to 50mM Tris buffer at pH 10 with distilled water. 1M Tris buffer were made, pH were adjusted to pH 10 with the addition of 1M HCl as stock solution stored in fridge. 500mM Tris buffer at pH 10 were made by diluting from 1M Tris buffer. Please refer to [[User:Melissa Novy/Notebook/CHEM-571/2012/09/12|Melissa's Notebook]] for details on the calculation on the actual and theoretical mass and concentration obtained for 100mM Tris buffer.
 +
* Absorbency were measured using UV-vis spectrometer for the sets of Au/BSA samples made on [[User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/09/11|2012/09/11]]. 1mL of each sample were used for measurement. For details of the absorbance, see [[User:Dhea Patel/Notebook/Experimental Biological Chemistry Notebook/2012/09/12|Dhea's Notebook]] for the absorbance obtained for Au/BSA sample after 4 hours of incubation.
 +
* 4mL of 133 Au/BSA solution made on 09/11/12 were mixed with 1mL of 50mM Tris buffer at pH 10, yielding a total of 10mM tris buffer in solution. 4mL of 136 Au/BSA solution were mixed with 1mL of 100mM Tris buffer at pH 10, yielding a total of 20mM tris buffer in solution. And 4mL of 138 Au/BSA solution were mixed with 1mL of 500M Tris buffer at pH 10, yielding a total of 100mM tris buffer in solution. All three samples were lightly shaken and let to re-suspend for 30 minutes. Absorbance for the three samples were taken after 30 minutes and after 90 minutes. See [[User:Dhea Patel/Notebook/Experimental Biological Chemistry Notebook/2012/09/12|Dhea's Notebook]] for results on the absorbance versus wavelength data on each Au/BSA sample. It was concluded that none of the solution were resuspended with the purple fibrous pellet, and the previous experiments cannot be repeated. Tris buffer mixture with Au/BSA resolution remained inconclusive.
 +
==Notes==
 +
* New sets of solutions were taken out of the incubator. Fibers were formed in all samples. It was suspected that the ratios prepared were wrong and all ratios are predicted to be above 133 Au/BSA. Supernatant were clear and fibers were formed closely together as pellet.
 +
* Sets of solutions made on [[User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/09/11|2012/09/11]] has produced homogeneous solutions at lower ratio and fibers at higher ratios of Au/BSA as previous experiments performed by other researchers. It was concluded that the ratios of Au/BSA calculations were correct, and that the procedure were reproducible. For details of the solution, please refer to [[User:Dhea Patel/Notebook/Experimental Biological Chemistry Notebook/2012/09/12|Dhea's notebook]] for the appearance of solution for a better understanding of the structural formation of fibers and appearance of pellet and supernatant after incubation.
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Purpose

  • To check if solutions made on 2012/09/11 has produced fibers
  • To mix supernatant with pellet in each sample with the addition of different concentration of Tris buffer at pH 10 and check re-suspension

Procedure

  • Sets of Au/BSA solutions with different ratios were remade from ratio 80 to ratio 170. For details with the volume, see Dhea's Notebook. The sets of solutions were incubated in 85°C for four hours. After four hours of incubation, 1mL of supernatant were measured for absorption activity under UV-vis from 700nm to 200nm. See Dhea's Notebook for details on absorption of each Au/BSA solutions after 4 hours of incubation.
  • 100mM Tris buffer at pH 10 were diluted to 50mM Tris buffer at pH 10 with distilled water. 1M Tris buffer were made, pH were adjusted to pH 10 with the addition of 1M HCl as stock solution stored in fridge. 500mM Tris buffer at pH 10 were made by diluting from 1M Tris buffer. Please refer to Melissa's Notebook for details on the calculation on the actual and theoretical mass and concentration obtained for 100mM Tris buffer.
  • Absorbency were measured using UV-vis spectrometer for the sets of Au/BSA samples made on 2012/09/11. 1mL of each sample were used for measurement. For details of the absorbance, see Dhea's Notebook for the absorbance obtained for Au/BSA sample after 4 hours of incubation.
  • 4mL of 133 Au/BSA solution made on 09/11/12 were mixed with 1mL of 50mM Tris buffer at pH 10, yielding a total of 10mM tris buffer in solution. 4mL of 136 Au/BSA solution were mixed with 1mL of 100mM Tris buffer at pH 10, yielding a total of 20mM tris buffer in solution. And 4mL of 138 Au/BSA solution were mixed with 1mL of 500M Tris buffer at pH 10, yielding a total of 100mM tris buffer in solution. All three samples were lightly shaken and let to re-suspend for 30 minutes. Absorbance for the three samples were taken after 30 minutes and after 90 minutes. See Dhea's Notebook for results on the absorbance versus wavelength data on each Au/BSA sample. It was concluded that none of the solution were resuspended with the purple fibrous pellet, and the previous experiments cannot be repeated. Tris buffer mixture with Au/BSA resolution remained inconclusive.

Notes

  • New sets of solutions were taken out of the incubator. Fibers were formed in all samples. It was suspected that the ratios prepared were wrong and all ratios are predicted to be above 133 Au/BSA. Supernatant were clear and fibers were formed closely together as pellet.
  • Sets of solutions made on 2012/09/11 has produced homogeneous solutions at lower ratio and fibers at higher ratios of Au/BSA as previous experiments performed by other researchers. It was concluded that the ratios of Au/BSA calculations were correct, and that the procedure were reproducible. For details of the solution, please refer to Dhea's notebook for the appearance of solution for a better understanding of the structural formation of fibers and appearance of pellet and supernatant after incubation.


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