Human and Mouse Apoptosis regulator Bcl-2 TranslationBlockerâ¢ siRNA Duplex Product Attributes
Blocks transcription and translation of Apoptosis regulator Bcl-2 from: Human and Mouse.
May also react with Apoptosis regulator Bcl-2 based upon sequence homology and clone specific publications.
Type of RNAi: siRNA.
siRNA target sequence: NM_000657.
Samples and cell lines tested: Hct116 Colorectal Carcinoma Cells.
Buffer and Stabilizer: Lyophilized. See datasheet for resuspension instructions and buffer information.
Concentration of Anti-Human and Mouse Apoptosis regulator Bcl-2: 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.
Apoptosis regulator Bcl-2 Knockdown Protocol:
- Starting with cultured cells in growth media seed 6 well plates with 1.5x105 cells and incubate at 37°C for 24 hours.
- Prepare transfection reagent in serum-free growth media such as Opti-MEM allow to come to room temperature. The above data was generated with Oligofectamine for delivery of human / mouse Bcl-2 siRNA prepared in accordance with manufacturerÂs instructions.
- Dilute siRNA to working concentration in serum-free growth media. For Oligofectamine or similar transfections into 6 well plates, we recommend starting siRNA concentration optimization at around 100 pmol siRNA duplex in 750 ul of serum free media. Optimal dilution should be experimentally determined.
- Combine diluted siRNA and transfection reagent. Equivalent volumes of prepared Oligofectamine or other transfection reagent and diluted siRNA should be combined and incubated at room temperature for 30 minutes with gentle mixing (optional).
- Add prepared siRNA solution to cells. Additionally, FITC labeled control siRNA (Product Number QC-1F) or positive control siRNA (Beta-Actin; Product Number QC-2) may be used to assess transfection efficiency.
- 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. Optimal knockdown time should also be experimentally determined. Optionally, a second transfection can be performed 24 hours after the first transfection to enhance knockdown. Human Bcl-2 knockdown at the protein level was measured at 24, 48, and 72 hours post transfection via western blot.
Apoptosis regulator Bcl-2 General Information
Gene Name: BCL2
Entrez Gene ID: 596
NCBI Ref Seq Accession Number: NP_000648
Apoptosis regulator Bcl-2 Molecular Weight:
Alternate Names: Bcl-2, PPP1R50
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.