Human ErbB4 / Her4 TranslationBlockerâ¢ siRNA Duplex Product Attributes
Blocks transcription and translation of ErbB4 / Her4 from: Human.
May also react with ErbB4 / Her4 based upon sequence homology and clone specific publications.
Type of RNAi: siRNA.
siRNA target sequence: NM_005235.
Samples and cell lines tested: T-47d Human Carcinoma Cells from Breast.
Buffer and Stabilizer: Lyophilized. See datasheet for resuspension instructions and buffer information.
Concentration of Anti-Human ErbB4 / Her4: 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.
ErbB4 / Her4 Knockdown Protocol:
- Starting with cultured cells in growth media seed 12 well plates with 5x104 cells and incubate at 37°C.
- Prepare transfection reagent in serum-free growth medium such as Opti-MEM-I. Allow to come to room temperature. Oligofectamine was used to generate the above data in accordance with manufacturerÂs instructions.
- Dilute siRNA to working concentration in serum-free growth media. For Oligofectamine or similar transfections into 12 well plates, we recommend starting siRNA concentration optimization at around 100-150 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 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. For 12 well plates this will entail adding the solution to cells in approximately 1.1 ml serum-free medium. 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 an initial 4 hour incubation 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. Alternatively, the media may be directly supplemented with serum such as 10% FBS.
- 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. ErbB4 / Her4 knockdown at the protein level was measured at 72 hours post transfection via western blot.
ErbB4 / Her4 General Information
Gene Name: ERBB4
Entrez Gene ID: 2066
NCBI Ref Seq Accession Number: NP_005226
ErbB4 / Her4 Molecular Weight:
Alternate Names: p180erbB4, HER4, ALS19, Erb-b4, ErbB-4
- Pan, J., Huang, H., Sun, L., Fang, B. & Yeung, S.-C. J. Bcl-2-associated X protein is the main mediator of manumycin a-induced apoptosis in anaplastic thyroid cancer cells. J. Clin. Endocrinol. Metab. 90, 3583Â91 (2005).
- BidÃ¨re, N. et al. Cathepsin D triggers Bax activation, resulting in selective apoptosis-inducing factor (AIF) relocation in T lymphocytes entering the early commitment phase to apoptosis. J. Biol. Chem. 278, 31401Â11 (2003).
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.