Human ErbB2 / HER2 TranslationBlockerâ¢ siRNA Duplex Product Attributes
Blocks transcription and translation of ErbB2 / HER2 from: Human.
May also react with ErbB2 / HER2 based upon sequence homology and clone specific publications.
Type of RNAi: siRNA.
siRNA target sequence: NM_004448.
Samples and cell lines tested: BT-474 Human Breast Carcinoma Cells, SkBr3 Human Breast Adenocarcinoma Cells, and MCF7 Human Breast Adenocarcinoma Cells.
Buffer and Stabilizer: Lyophilized. See datasheet for resuspension instructions and buffer information.
Concentration of Anti-Human ErbB2 / HER2: 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.
ErbB2 / HER2 Knockdown Protocol:
- Starting with cultured cells in growth media seed 6 well plates with 3.5x105 cells and incubate at 37°C.
- Prepare transfection reagent in serum-free growth media such as Opti-MEM-I. Allow to come to room temperature. The above data was generated with TransIT-TKO for delivery of ErbB2 / Her2 siRNA prepared in accordance with manufacturerÂs instructions.
- Dilute siRNA to working concentration in serum-free growth media. For TransIT-TKO or similar transfections into 6 well plates, we recommend starting siRNA concentration optimization at around 75-100 pmol siRNA duplex in 200 ul of serum free media.Optimal dilution should be experimentally determined.
- Combine diluted siRNA and transfection reagent. Equivalent volumes of prepared TransIT-TKO 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. For 6 well plates this will entail adding the solution to cells in approximately 2.5 ml serum-free media. 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. The above data was generated with a 18 hour initial incubation.
- Wash cells and add complete growth 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. ErbB2 / Her2 knockdown at the protein level was measured at 72 hours post transfection via western blot.
ErbB2 / HER2 General Information
Gene Name: ERBB2
Entrez Gene ID: 2064
NCBI Ref Seq Accession Number: NP_004439
ErbB2 / HER2 Molecular Weight:
Alternate Names: CD340, NGL, HER-2/neu, HER2, MLN 19, NEU, HER-2, TKR1, Receptor tyrosine-protein kinase erbB-2
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.