Objective
- Setup AgNP and AuNP synthesis reactions for SPI, WPI, Casein, Pea Protein and Brown Rice Protein
- Further Research reducing sugars and other extra-protein reducing agents
Materials
- Used same nutritional proteins on 02/02/16
- Protein and gold stocks prepared fresh every day
Procedure
Reaction procedure devised on 02/03/16 prepared and executed today.
- Reducing stock protein concentrations (and thus concentrations in reaction mix) to increase metal/protein ratio to spur NP formation as we saw protein aggregate and nanofiber formation in previous reactions
- Hold protein volume and thus concentration) constant and change fructose concentration (reducing agent)
Detailed Reaction Setup for 02/09/16 Shown Below
1mL Reactions
|
'
|
'
|
'
|
'
|
'
|
Note that many water volumes were too high, and therefore reactions were more dilute than antipated
|
'
|
'
|
|
Tube # |
2.5mM HAuCl4 (µL) |
2.5mMAgNO3 (µL) |
Protein (µL) |
5mM Fructose (µL) |
Water (µL) |
Au/Ag:Protein |
Notes
|
|
1 |
40 |
0 |
560 |
0 |
400 |
|
No change
|
2mg/mL Casein |
2 |
40 |
0 |
560 |
50 |
350 |
|
|
grams weighed: 20mg |
3 |
40 |
0 |
560 |
100 |
300 |
|
|
Stock: 10mL |
4 |
0 |
40 |
560 |
0 |
400 |
|
No change
|
|
5 |
0 |
40 |
360 |
0 |
600 |
|
|
|
6 |
0 |
40 |
200 |
0 |
760 |
|
|
|
7 |
40 |
0 |
560 |
0 |
400 |
|
pink aggregate
|
|
8 |
40 |
0 |
560 |
100 |
300 |
|
|
|
9 |
40 |
0 |
440 |
0 |
520 |
|
darker pink/purple aggregate (maybe fibers)
|
0.75mg/mL SPI |
10 |
40 |
0 |
440 |
100 |
420 |
|
|
grams weighed: 7.5mg |
11 |
40 |
0 |
280 |
0 |
680 |
|
dark pink/purple fibers
|
Stock: 10mL |
12 |
40 |
0 |
280 |
100 |
580 |
|
|
|
13 |
0 |
40 |
560 |
0 |
400 |
|
white aggregate at bottom of tube
|
|
14 |
0 |
40 |
560 |
100 |
300 |
|
|
|
15 |
0 |
40 |
440 |
0 |
520 |
|
|
|
16 |
0 |
40 |
440 |
100 |
420 |
|
|
|
17 |
0 |
40 |
280 |
0 |
680 |
|
|
|
18 |
0 |
40 |
280 |
100 |
580 |
|
|
|
19 |
40 |
0 |
380 |
0 |
620 |
|
19-20 White aggregates at bottom of tube and sides
|
|
20 |
40 |
0 |
360 |
0 |
640 |
|
|
|
21 |
40 |
0 |
340 |
0 |
660 |
|
21, slight pinkish tinge to white aggregates
|
1mg/mL Whey Protein |
22 |
0 |
40 |
560 |
0 |
400 |
|
No change
|
grams weighed: 10mg |
23 |
0 |
40 |
560 |
100 |
300 |
|
|
Stock: 10mL |
24 |
0 |
40 |
360 |
0 |
600 |
|
|
|
25 |
0 |
40 |
360 |
100 |
500 |
|
|
|
26 |
0 |
40 |
200 |
0 |
760 |
|
|
|
27 |
0 |
40 |
200 |
100 |
660 |
|
|
|
28 |
40 |
0 |
360 |
0 |
640 |
|
Dark purple specks at bottom of the tube (not much on walls)
|
|
29 |
40 |
0 |
360 |
50 |
590 |
|
Lower protein concentration, as each lower volume of protein added, created darker and more particles
|
|
30 |
40 |
0 |
360 |
100 |
540 |
|
|
0.5mg/mL Pea Protein |
31 |
0 |
40 |
560 |
0 |
400 |
|
white aggregates at bottom of tube only
|
grams weighed: 10mg |
32 |
0 |
40 |
560 |
100 |
300 |
|
|
Stock: 10mL |
33 |
0 |
40 |
440 |
0 |
520 |
|
|
|
34 |
0 |
40 |
440 |
100 |
420 |
|
|
|
35 |
0 |
40 |
360 |
0 |
600 |
|
|
|
36 |
0 |
40 |
360 |
100 |
500 |
|
|
|
37 |
40 |
0 |
360 |
0 |
640 |
|
dark purple specks on bottom and sides of tube (mostly on bottom)
|
|
38 |
40 |
0 |
360 |
50 |
590 |
|
- change protein to metal ratios, maybe change fructose to citrate
|
1mg/mL Rice Protein |
39 |
40 |
0 |
360 |
100 |
540 |
|
|
grams weighed: 10mg |
40 |
0 |
40 |
560 |
0 |
400 |
|
white aggregates at bottom of tube
|
Stock: 10mL |
41 |
0 |
40 |
560 |
100 |
300 |
|
|
|
42 |
0 |
40 |
440 |
0 |
520 |
|
|
|
43 |
0 |
40 |
440 |
100 |
420 |
|
|
|
44 |
0 |
40 |
360 |
0 |
600 |
|
|
|
45 |
0 |
40 |
360 |
100 |
500 |
|
|
|
Results
No reactions worked, this was due to some errors. Some water volumes were calculated higher than 1ml (all 1ml reactions). And thus everything in reaction mix was more dilute than necessary. Although the max this occurred at was 1.05ml/1.00ml.
No other reactions worked. Protein concentration was too high for casein (which was tested at 2mg/ml).
|