DISCONTINUED: Human Insulin Receptor TranslationBlockerâ¢ siRNA Duplex Product Attributes
Blocks transcription and translation of Insulin Receptor from: Human.
May also react with Insulin Receptor based upon sequence homology and clone specific publications.
Type of RNAi: siRNA.
siRNA target sequence: NM_000208.
Samples and cell lines tested: HepG2 Human Hepatocellular Carcinoma Cells.
Buffer and Stabilizer: Lyophilized. See datasheet for resuspension instructions and buffer information.
Concentration of Anti-Human Insulin Receptor: 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.Insulin Receptor Knockdown Protocol:
- Starting with cultured cells in growth media seed plates with cells and incubated at 37Â°C. For instance, seed 6 well plates with 2x105 cells in growth media and incubate at 37Â°C until cells reach 40% confluence. Transfection reagents may specify the desired level of confluence to look for prior to starting transfections.
- The above data was generated with N-TER nanoparticle transfection agent in accordance with manufacturerÂs instructions. Prepare transfection reagent in antibiotic-free growth medium. For other transfection reagents, serum-free media may be required. Allow to come to room temperature. </ li>
- Dilute siRNA to working concentration in antibiotic-free growth media. For N-TER or similar transfections into 6 well plates, we recommend starting siRNA dose optimization at approximately 75-100 pmol siRNA duplex.Optimal dilution should be experimentally determined.
- Combine diluted siRNA and transfection reagent according to manufacturerÂs instructions. Ratio of transfection reagent to siRNA should be optimized.
- 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 medium. Optionally, 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 and assay well 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. Insulin Receptor knockdown at the protein level was measured at 72 hours post transfection via Western Blot.
Insulin Receptor General Information
Gene Name: Insr
Entrez Gene ID: 3643
NCBI Ref Seq Accession Number: NP_000199
Insulin Receptor Molecular Weight:
Alternate Names: InsR, IR, CD220
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.
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