Human Alpha-synuclein TranslationBlockerâ¢ siRNA Duplex Product Attributes
Blocks transcription and translation of Alpha-synuclein from: Human.
May also react with Alpha-synuclein based upon sequence homology and clone specific publications.
Type of RNAi: siRNA.
siRNA target sequence: NM_000345.
Samples and cell lines tested: SH-SY5Y.
Buffer and Stabilizer: Lyophilized. See datasheet for resuspension instructions and buffer information.
Concentration of Anti-Human Alpha-synuclein: Lyophilized
SiRNA Purification Method:RPC/HPLC
Storage Conditions: Store Lyophilized siRNA duplex at 4C. Reconsituted product may be stored at -20C for up to 1 month.
Alpha-synuclein Knockdown Protocol:
- Starting with cultured cells in growth media (+10% fetal calf serum (FCS)) seed 6 well plates with 3 x 105 cells and incubate at 37°C for 24 hours.
- Prepare transfection reagent in serum-free growth media (Opti-MEM used in this experiment) and allow to come to room temperature. The above data was generated with Lipofectamine 2000 diluted 1:100 prepared in accordance with manufacturerÂs instructions.
- Dilute siRNA to working concentration in serum-free growth media. The above data was generated with 60-120 pmol of QX2 siRNA duplex in 300 ul of media. Optimal dilution should be experimentally determined.
- Combine diluted siRNA and transfection reagent. In this case, 100 ul of Lipofectamine 2000 was incubated with 100 ul of diluted siRNA for 30 minutes with gentle mixing (optional).
- Add prepared siRNA solution to cells.
- Incubate for several hours in serum free media. In the above experiment, a 6 hour incubation was performed.
- Add serum containing media to culture. Data above was generated by adding 1.5x culture volume of media containing 30% FBS (2% final concentration).
- Incubate and assay several hours after transfection. An additional transfection may be performed, as above, 24 hours after initial transfection to enhance knockdown. Optimal knockdown time should also be experimentally determined. In the above experimental conditions, SNCA knockdown at the protein level was confirmed 72 hours post transfection via western blot.
Alpha-synuclein General Information
Gene Name: SNCA
Entrez Gene ID: 6622
NCBI Ref Seq Accession Number: NP_000336
Alpha-synuclein Molecular Weight:
Alternate Names: PD1, NACP, PARK1, PARK4, SNCA
Limitations and Warranty
This product is for Research Use Only. This product is guaranteed to work for a period of two years when stored at -70C or colder, and one year when aliquoted and stored at -20C.