| Cell Line Species/Tissue: |
Human
/
neuroblastoma |
| Transfection Reagent: |
RNAiFect |
Nucleic Acid:
|
siRNA (dsRNA) |
Growth Medium:
|
Iscove‘s modified Dulbecco‘s medium (IMDM; Seromed) supplemented with 10% FCS, 100 IU/ml penicillin, and 100 j.tg/mi streptomycin at 37 °C in a humidified 5% CO2 incubator. |
Percent Serum (%):
|
5% FCS |
Reporter System:
|
|
Plasmid Purification Method:
|
|
Plate Format:
|
no information |
Number of Cells:
|
|
Percent Confluence(%):
|
70-80% |
Amount of Nucleic Acid (µg):
|
7 nM |
Amount of Enhancer (µl):
|
|
Amount of Reagent (µl):
|
no information |
Complex Incubation on Cells (hrs):
|
|
Analysis Performed Post-Transfection (hrs):
|
48 h |
Transfection Efficiency (%):
|
|
| Knockdown Efficiency (%): |
no information |
| |
| Any modifications to the protocol?:
|
| |
| Notes: |
| siRNA was constructed directed against two different regions of ST8SiaIV (PST 1+11). PST 1: 5‘ GGCUGGCUCUUCAAUCUUtt 3‘ and 5‘ GAA GAUUGAAGAGCCAGCCt
3‘; PST II: 5‘ GGUUGGAAAAUCAAUUCCUtt 3‘ and 5‘ AGGAAUUGAUUUUCCAACCtt 3‘.
siRNA/RNAiFect ratio of 1:6
To analyze the adhesion capacity of transfected cells, cells were collected 48 h later, and 0.5x 106 NB cells were then added to the HUVEC rnonolayer for 60 min.
|
| References: |
| ELSEVIER
International Immunopharmacology 5 (2005) 757—769
Valproic acid modulates NCAM polysialylation and polysialyltransferase mRNA expression in human tumor cells
Wolf-Dietrich Beecken, Tobias Engl, Henry Ogbomo, Boma Reija, Jindrich Cinatl,
Jürgen Bereiter-Hahn, Elsie Oppermann, Dietger Jonas, Roman A. Blaheta*
|