User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/11/20
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==Purpose== | ==Purpose== | ||
| + | * ADA protein were transferred from dialysis tubing into 15mL falcon tubes | ||
* Make Au/ADA samples with the following mole ratios: 60 - 70 - 80 - 90 - 100 - 110 - 120 - 130 - 140 - 150, with ADA fraction 2 after dialysis. | * Make Au/ADA samples with the following mole ratios: 60 - 70 - 80 - 90 - 100 - 110 - 120 - 130 - 140 - 150, with ADA fraction 2 after dialysis. | ||
| - | * | + | * Un-dialyzed Au/ADA samples and Au/HRP made on [[User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/11/14|2012/11/14]] were run on UV-vis spectrometer and Atomic Absorption Spectrometer in order to compare spectra results for Au/ADA, Au/HRP, Au/Lysozyme, and Au/BSA. |
| - | * | + | * Resuspend Au/HRP samples in different concentration and pH of Tris buffer to test ionic strength. |
| + | ==Procedure for Dialysis== | ||
| + | * Dialysis beaker containing dialysis tubing enclosed with ADA protein fractions were taken from the 4°C cold room into room temperature lab room. | ||
| + | * The dialysis clips were taken off from dialysis tubing. ADA protein fractions were poured into a sterile 15mL falcon tubes. | ||
| + | * This process was repeated for all three protein fractions: ADA fraction 1&3 made on [[User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/11/06|2012/11/06]], ADA fraction 2 made on [[User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/11/06|2012/11/06]], and ADA fraction purified on [[User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/10/03|2012/10/03]]. | ||
| + | * ADA fractions in 15mL falcon tubes were stored in 4°C refrigerator. | ||
| + | ==Procedure for making dialyzed Au/ADA samples== | ||
| + | * Au/ADA samples were made with the following mole ratios: | ||
| + | 60 - 70 - 80 - 90 - 100 - 110 - 120 - 130 - 140 - 150 | ||
| + | * Stock solution of HAuCl<sub>4</sub> was made on [[User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/09/05|2012/09/05]] with a concentration of 10.5uM. | ||
| + | * Stock solution for ADA was the ADA protein fraction made in [[User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/10/03|2012/10/03]] after dialysis. The ADA stock solution has a concentration of 58.36μM. | ||
| + | * Volume of ADA protein was set at 137.1uL and a range of HAuCl<sub>4</sub> was used from 45.71μL to 114.3μL. Water was added to the sample to increase the volume of sample to 8mL. Volumes of each reactants are shown in table below: | ||
| + | {| {{table}} | ||
| + | | align="center" style="background:#f0f0f0;"|'''Au/ADA ratio''' | ||
| + | | align="center" style="background:#f0f0f0;"|'''ADA added[uL]''' | ||
| + | | align="center" style="background:#f0f0f0;"|'''HAuCl4 Added [uL]''' | ||
| + | | align="center" style="background:#f0f0f0;"|'''Water Added[uL]''' | ||
| + | | align="center" style="background:#f0f0f0;"|'''[ADA]final[uM]''' | ||
| + | | align="center" style="background:#f0f0f0;"|'''[HAuCl4]final[uM]''' | ||
| + | |- | ||
| + | | 60||137.1||45.71||7817.2||1||60 | ||
| + | |- | ||
| + | | 70||137.1||53.3||7809.6||1||70 | ||
| + | |- | ||
| + | | 80||137.1||60.9||7802||1||80 | ||
| + | |- | ||
| + | | 90||137.1||68.6||7794.4||1||90 | ||
| + | |- | ||
| + | | 100||137.1||76.2||7786.8||1||100 | ||
| + | |- | ||
| + | | 110||137.1||83.8||7779.1||1||110 | ||
| + | |- | ||
| + | | 120||137.1||91.4||7771.5||1||120 | ||
| + | |- | ||
| + | | 130||137.1||99||7763.9||1||130 | ||
| + | |- | ||
| + | | 140||137.1||106.7||7756.3||1||140 | ||
| + | |- | ||
| + | | 150||137.1||114.3||7748.7||1||150 | ||
| + | |- | ||
| + | | | ||
| + | |} | ||
| + | |||
| + | * The samples were made in 15mL non-sterile falcon tubes. After all reactants were added, samples are capped and wrapped around with aluminum foil. | ||
| + | * Samples were placed in incubator at 85°C for 4 hours then cooled down to room temperature. | ||
| + | |||
| + | ==Procedure for Running UV-vis spectrometer and Atomic Absorption spectrometer on Au/ADA and Au/HRP samples== | ||
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Revision as of 15:14, 27 November 2012
 Experimental Biological Chemistry I
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Purpose
Procedure for Dialysis
Procedure for making dialyzed Au/ADA samples
60 - 70 - 80 - 90 - 100 - 110 - 120 - 130 - 140 - 150
Procedure for Running UV-vis spectrometer and Atomic Absorption spectrometer on Au/ADA and Au/HRP samples | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
