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

Purpose

• To check if solutions made on 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
• Abigail E. Miller 10:55, 17 September 2012 (EDT):I'm confused by this. are you adding tris to the pelletonly, to just the supernatant or to both?
• Prepare the ratio 80 Au/BSA sample for peak shift testing next week
• Abigail E. Miller 10:55, 17 September 2012 (EDT):explain what hte peak shift experiemnt is.

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. For details of the results, see Dhea's Notebook.
• Abigail E. Miller 10:58, 17 September 2012 (EDT):masses and volumes and ratios. can reference the calculations and precise details are in Dhea's notebook.
• 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.
• Abigail E. Miller 10:58, 17 September 2012 (EDT):masses and volumes and ratios. can reference the calculations and rpecise details - theoretical mass, acutal mass, etc..
• 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.
• Abigail E. Miller 10:58, 17 September 2012 (EDT): show data here too.
• 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.
• Abigail E. Miller 10:58, 17 September 2012 (EDT):show data here too. conclusions form data?

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 according to predicted.
• Abigail E. Miller 10:59, 17 September 2012 (EDT):be specific. what was predicted? photos of solutions? what ratios produced fibers? what produced nothing? what produced homogenous solutions?