User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/10/31

(Difference between revisions)

Revision as of 23:44, 7 December 2012

Experimental Biological Chemistry I

Au/Lysozyme samples were made with the same procedure used to make Au/BSA solutions.

Lysozyme in powder form from chicken egg white obtained from Signma® is used to make the lysozyme stock solution.
5mL of 70μM lysozyme stock solution was made in the following way:
- 0.005g of lysozyme was weighted out
- 0.005g of lysozyme was transferred to a 15ml falcon tube and mixed well with 5mL of distilled water.
- See Melissa's Notebook for calculation that stated that 0.005g of lysozyme was needed for 5mL of 70μM of lysozyme stock solution.
The HAuCl₄ used to make Au/Lysozyme samples was already made on 2012/09/05, and the concentration of the HAuCl₄ was 10mM.
Au/Lysozyme samples was made with mole ratios listed below:

  20 - 30 - 40 - 50 - 60 - 70 - 80 - 100 - 120 - 130

Based off the calculations for Au/BSA samples, a few changes were made to calculate the amount of lysozyme and HAuCl₄ needed for Au/Lysozyme samples.
The table below listed the initial conditions for both lysozyme and HAuCl₄ both theoretically and actually:

-	Lysozyme	HAuCl₄
Molecular Weight [g/mol]	14300	339.79
Theoretical Concentration [M]	7.00E-05	0.01
Volume of Water to add [L]	0.005	0.025
Moles Present [mol]	3.50E-07	0.00025
Theoretical Mass Needed [g]	0.005	0.0849
Actual Mass Weighted [g]	0.005	0.0892
Actual Concentration [M]	0.0007	10501

With the above calculation, the actual concentration of lysozyme came out to be 70μM and the actual concentration of HAuCl₄ came out to be 10.501mM. The two concentration was used to calculate the amount of water, lysozyme, HAuCl₄ needed to make the ratios listed above.
The table below listed the amount of water, lysozyme, HAuCl₄ used for each Au/Lysozyme sample:

Au/Lysozyme ratio	Lysozyme added [uL]	HAuCl4 added [uL]	Water added [uL]	Final Concentration of Au[mM]	Final Concentration of Lysozyme[uM]
20	114.3	15.2	7870.5	1	20
30	114.3	22.9	7862.8	1	30
40	114.3	30.5	7855.2	1	40
50	114.3	38.1	7847.6	1	50
60	114.3	45.7	7840	1	60
70	114.3	53.3	7832.4	1	70
80	114.3	60.9	7824.8	1	80
100	114.3	76.2	7809.5	1	100
120	114.3	91.4	7794.3	1	120
130	114.3	99	7786.7	1	130

The stock solutions were added with pipetman and non-sterile pipette tips. All samples were made in 10mL large test tubes. All samples were capped and wrapped in aluminum foil.
All samples were placed in incubator at 85°C for four hours then in room temperature to cool down.

@@ Line 47: / Line 47: @@
 | align="center" style="background:#f0f0f0;"|'''HAuCl4 added [uL]'''
 | align="center" style="background:#f0f0f0;"|'''Water added [uL]'''
+| align="center" style="background:#f0f0f0;"|'''Final Concentration of Au[mM]'''
+| align="center" style="background:#f0f0f0;"|'''Final Concentration of Lysozyme[uM]'''
 |-
-| 20||114.3||15.2||7870.5
+| 20||114.3||15.2||7870.5||1||20
 |-
-| 30||114.3||22.9||7862.8
+| 30||114.3||22.9||7862.8||1||30
 |-
-| 40||114.3||30.5||7855.2
+| 40||114.3||30.5||7855.2||1||40
 |-
-| 50||114.3||38.1||7847.6
+| 50||114.3||38.1||7847.6||1||50
 |-
-| 60||114.3||45.7||7840
+| 60||114.3||45.7||7840||1||60
 |-
-| 70||114.3||53.3||7832.4
+| 70||114.3||53.3||7832.4||1||70
 |-
-| 80||114.3||60.9||7824.8
+| 80||114.3||60.9||7824.8||1||80
 |-
-| 100||114.3||76.2||7809.5
+| 100||114.3||76.2||7809.5||1||100
 |-
-| 120||114.3||91.4||7794.3
+| 120||114.3||91.4||7794.3||1||120
 |-
-| 130||114.3||99||7786.7
+| 130||114.3||99||7786.7||1||130
 |}