Human ApoE TranslationBlockerâ¢ siRNA Duplex Product Attributes
Blocks transcription and translation of ApoE from: Human.
May also react with ApoE based upon sequence homology and clone specific publications.
Type of RNAi: siRNA.
siRNA target sequence: NM_000041.
Samples and cell lines tested: Ovcar3 Human Ovarian Cancer Cells.
Buffer and Stabilizer: Lyophilized. See datasheet for resuspension instructions and buffer information.
Concentration of Anti-Human ApoE: 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.
ApoE Knockdown Protocol:
- Starting with cultured cells in growth media seed 24 well plates with 5 x 104 cells and incubate at 37°C for 24 hours.
- Prepare transfection reagent in serum-free growth media allow to come to room temperature. The above data was generated with Oligofectamine prepared in accordance with manufacturerÂs instructions.
- Dilute siRNA to working concentration in serum-free growth media. Optimal dilution should be experimentally determined.
- Combine diluted siRNA and transfection reagent. Equivalent volumes of prepared Oligofectamine and diluted siRNA should be combined and incubated at room temperature for 20 minutes with gentle mixing (optional).
- Add prepared siRNA solution to cells.
- Incubate for several hours in serum free media.
- Add serum containing media to culture without a media change. Generally 1-2 fold of the original volume of siRNA containing serum free media is added to restore nutrient levels in media.
- 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, Apolipoprotein E (ApoE) knockdown at the protein level was not visible until 24 hours post transfection. Knockdown continued to improve
ApoE General Information
Gene Name: APOE
Entrez Gene ID: 348
NCBI Ref Seq Accession Number: NP_000032
ApoE Molecular Weight:
Alternate Names: AD2, APO-E, LPG, LDLCQ5, ApoE4, ApoE, Apolipoprotein E
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.