06/19/13
- Luc assay, time point 2
- PcTF transfection for U2OS, outgrowth #2
Luciferase activity assay - time point 2
Cell prep
- Harvested cells
- Seeded new dox+ plate with 1/2 of cells
- Pelleted other half, resuspended in 2 mL FACS buffer
Assay reagents
- Used the Biotium Steady-Luc Firefly HTS Assay Kit (same as 6/18/13)
Luc assay
- Filtered 700 μL cells through strainer caps
- Other steps same as 6/18/13
Cell counts
- Wang lab's Accuri flow cytometer.
- Set machine to read 20 uL of cells
- Be sure to "clean" with water-run in between samples
Sample ID |
Gated count/ 20 μL |
|
Cells/ 100 μL
|
sample 1 |
10,880 |
x5 = |
54,400
|
sample 2 |
10,430 |
x5 = |
52,150
|
sample 3 |
11,897 |
x5 = |
59,485
|
sample 4 |
10,316 |
x5 = |
51,580
|
sample 5 |
9,784 |
x5 = |
48,920
|
sample 6 |
11,549 |
x5 = |
57,745
|
PcTF transfection for U2OS, outgrowth #2
- Expanded transfected and mock-transfected cells from old 10 cm plates (6/15/13)
- 1:10 passage of PcTF+ or mock cells into three new 10 cm plates labeled "A, B, C" (standard complete medium)
|