| Cell Line Species/Tissue: |
Rat
/
Primary Aortic endothelial cells |
| Transfection Reagent: |
HiPerFect |
Nucleic Acid:
|
siRNA (dsRNA) |
Growth Medium:
|
RPMI 1640 |
Percent Serum (%):
|
10% |
Reporter System:
|
|
Plasmid Purification Method:
|
|
Plate Format:
|
6-well |
Number of Cells:
|
1.5 x10e5 |
Percent Confluence(%):
|
50% |
Amount of Nucleic Acid (µg):
|
5nM |
Amount of Enhancer (µl):
|
|
Amount of Reagent (µl):
|
9µl |
Complex Incubation on Cells (hrs):
|
48-72h |
Analysis Performed Post-Transfection (hrs):
|
48-72h |
Transfection Efficiency (%):
|
|
| Knockdown Efficiency (%): |
>80% |
| |
| Any modifications to the protocol?:
|
| Addition of sRNA-HiPerFect complexes in cell suspension. See notes. |
| Notes: |
| Addition of sRNA-HiPerFect complexes in cell suspension, mixed by pipetting (very carefully) and then transferring of the cell suspension+siRNA to the plate.
Used the HiPerFect FastForward protocol (cell seeding and transfection at the same day). Checked cytotoxicity by using neutral red stating. No cytotoxicity could be observed. |
| References: |
| |