| Cell Line Species/Tissue: |
mouse
/
macrophage |
| Transfection Reagent: |
HiPerFect |
Nucleic Acid:
|
siRNA (dsRNA) |
Growth Medium:
|
DMEM |
Percent Serum (%):
|
10% |
Reporter System:
|
|
Plasmid Purification Method:
|
|
Plate Format:
|
24 well |
Number of Cells:
|
20.000 |
Percent Confluence(%):
|
|
Amount of Nucleic Acid (µg):
|
25nM |
Amount of Enhancer (µl):
|
|
Amount of Reagent (µl):
|
3µl |
Complex Incubation on Cells (hrs):
|
48 h |
Analysis Performed Post-Transfection (hrs):
|
48 h |
Transfection Efficiency (%):
|
|
| Knockdown Efficiency (%): |
75% |
| |
| Any modifications to the protocol?:
|
| Yes, see notes. |
| Notes: |
| Modification to the standard protocol:
1. Cell seeding (2x10e4 cells per well of a 24 well plate) in 100µl medium including serum
2. Complex formation in 100µl (medium without serum)
3. Add transfection complexes to cells and incubate for 6h
4. Add 400µl medium (containig serum) 6h after incubation. |
| References: |
| |