User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/11/07: Difference between revisions
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* Au/HRP samples were made with the mole ratios listed below: | * Au/HRP samples were made with the mole ratios listed below: | ||
10 - 50 - 100 - 150 - 200 - 250 - 300 - 350 - 400 - 450 | 10 - 50 - 100 - 150 - 200 - 250 - 300 - 350 - 400 - 450 | ||
* | * 28.5uM HRP stock solution was made. HRP from Alfa Aesar® has a molecular weight of 40,000 g/mol. 0.0057g of HRP was weighted out and mixed with 5mL of distilled water. The following calculations was done to obtain masses weighted for 28.5uM HRP stock solution: | ||
28.5uM HRP * 0.005L = 1.5E-7 mol | |||
1.5E-7mol * 40,000g/mol = 0.0057g of HRP | |||
* HAuCl<sub>4</sub> used for making Au/HRP samples were made on [[User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/09/05|09/05/2012]] and has a concentration of 10.5mM. | |||
* Volume of HRP was set at 280.7uL while volumes of HAuCl<sub>4</sub> were ranging from 7.62μL to 342.8μL. Distilled water was added until volume of sample at each mole ratio reached 8mL. Detailed volumes for each reactant is listed in table below: | |||
{| {{table}} | |||
| align="center" style="background:#f0f0f0;"|'''Au/HRP ratio''' | |||
| align="center" style="background:#f0f0f0;"|'''HRP added [uL]''' | |||
| align="center" style="background:#f0f0f0;"|'''HAuCl4 added [uL]''' | |||
| align="center" style="background:#f0f0f0;"|'''Water added [uL]''' | |||
|- | |||
| 10||280.7||7.62||7711.68 | |||
|- | |||
| 50||280.7||38.1||7681.2 | |||
|- | |||
| 100||280.7||76.2||7643.1 | |||
|- | |||
| 150||280.7||114.3||7605 | |||
|- | |||
| 200||280.7||152.4||7566.9 | |||
|- | |||
| 250||280.7||190.5||7528.8 | |||
|- | |||
| 300||280.7||228.5||7490.8 | |||
|- | |||
| 350||280.7||266.6||7452.7 | |||
|- | |||
| 400||280.7||304.7||7414.6 | |||
|- | |||
| 450||280.7||342.8||7376.5 | |||
|} | |||
* Each Au/HRP samples were capped, wrapped with aluminum foil, and placed in incubator at 85°C for 4 hours then cooled down to room temperature. | |||
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Revision as of 22:38, 27 November 2012
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Purpose
Procedure for Bradford Assay
Results and Calculations of ADA protein Concentrations
Absorbance = 27.609 * [BSA]mg/mL + 0.4363
Procedure for Making Au/HRP samples
10 - 50 - 100 - 150 - 200 - 250 - 300 - 350 - 400 - 450
28.5uM HRP * 0.005L = 1.5E-7 mol 1.5E-7mol * 40,000g/mol = 0.0057g of HRP
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