Experimental Goals
- Perform an Immunoprecipitation of iNOS/NOS2 from LPS +/- IFNg stimulated cell lysates.
- Preparation of 8% PAGE running gel.
Part One: IP of iNOS/NOS2
- Volumes of RIPA buffer required for IP. (See previous entry for calculation of volumes.)
- I am student A, Sunny is Student B, therefore the samples are divided as follows:
| Experimenter
|
Sample Number
|
Sample
|
Volume of sample required (µL)
|
Volume of RIPA buffer needed (µL)
|
| Eric |
1 |
24 hr (+) LPS |
64.9 |
285.1
|
| Eric |
2 |
48 hr (+) LPS |
74.7 |
275.3
|
| Eric |
3 |
72 hr (+) LPS |
97.9 |
252.1
|
| Eric |
4 |
24 hr (+) LPS (+) IFNg |
92.6 |
257.4
|
| Eric |
5 |
48 hr (+) LPS (+) IFNg |
128.8 |
221.2
|
| Eric |
6 |
72 hr (+) LPS (+) IFNg |
62.5 |
287.5
|
| Sunny |
7 |
24 hr (+) IFNg |
34.9 |
315.1
|
| Sunny |
8 |
48 hr (+) IFNg |
40.3 |
309.7
|
| Sunny |
9 |
72 hr (+) IFNg |
350.0 |
0.0
|
| Sunny |
10 |
24 hr control |
34.2 |
315.8
|
| Sunny |
11 |
48 hr control |
35.6 |
314.4
|
| Sunny |
12 |
72 hr control |
34.2 |
315.8
|
Part Two: 8% Acrylamide Gel
- Done with no problems encountered.
|
Project name
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