User:Karmella Haynes/Notebook/PcTF Genomics/2013/06/15: Difference between revisions
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== | ==06/15/13== | ||
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* | * Gal4-EED/luc Luciferase ICC - fixation, permeabilization, primary ab staining | ||
* Gal4-EED/ luc dox induction - shipment 2 cells | |||
* PcTF transfection for U2OS, outgrowth #1 | |||
---- | ---- | ||
''' | '''Gal4-EED/luc Luciferase ICC'''<br> | ||
Samples (Shipment 1 cells) | |||
# 0 μg/mL dox | |||
# 0.1 | |||
# 0.2 | |||
# 0.5 | |||
# 1.0 | |||
# 2.0 (induced on 6/02/13, 13 days) | |||
* Followed standard protocol | |||
* Harvested cells from one 6-well plate (split plate #2 for further analyses) | |||
* At the primary antibody staining step, keep samples and reagents on ice overnight in the cold room | |||
--> | ---- | ||
'''Gal4-EED/luc dox induction'''<br> | |||
Samples (Shipment 2 cells); new 6-well plate... | |||
# 0 μg/mL dox | |||
# 0.1 | |||
# 0.2 | |||
# 0.5 | |||
# 1.0 | |||
# 2.0 (induced on 6/02/13, 13 days) | |||
* For this set, test earlier time points and try luc activity assay | |||
---- | |||
'''PcTF transfection for U2OS, outgrowth #1''' | |||
* 6-well plates: 1-3 = +PcTF, 4-6 = mock (lipo only) | |||
* Expand cells from 6-well plates in individual 10 cm plates (10 mL medium). 6 10 cm plates total. | |||
* Use standard growth medium. | |||
* Use all cells. Do not dilute. | |||
Revision as of 20:11, 26 June 2013
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06/15/13
Gal4-EED/luc Luciferase ICC Samples (Shipment 1 cells)
Gal4-EED/luc dox induction Samples (Shipment 2 cells); new 6-well plate...
PcTF transfection for U2OS, outgrowth #1
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