Silencer™ siRNA Transfection II Kit
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Invitrogen™

Silencer™ siRNA Transfection II Kit

This Ambion™ kit is for the optimization of transfection protocols for specific cell types. It includes 0.4 mL of eachRead more
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Catalog number AM1631
Price (USD)
796.00
Each
Add to cart
Price (USD)
796.00
Each
Add to cart

This Ambion™ kit is for the optimization of transfection protocols for specific cell types. It includes 0.4 mL of each of two transfection agents; siPORT™ Amine Transfection Agent and siPORT™ NeoFX™ Transfection Agent along with control siRNAs.

  • Broad cell line compatibility-kit includes siPORT™ Amine and siPORT™ NeoFX™ transfection agents
  • Faster protocol optimization-kit includes ready-to-transfect GAPDH and negative siRNA controls

The choice of transfection agent is critical for gene silencing experiments. Every transfection agent has a different delivery efficiency when used with different cell types, and the overall transfection efficiency and degree of gene silencing depend on the nature of the transfection agent/siRNA complex. Whether you are new to RNAi research or you want to expand your experiments to include different cell lines, the Silencer™ siRNA Transfection II Kit enables you to optimize your transfection protocols for specific cell types. The Silencer™ siRNA Transfection II Kit includes siPORT™ Amine Transfection Agent (a polyamine mixture) and the new siPORT™ NeoFX™ Transfection Agent (a cationic and neutral lipid mixture). These reagents support efficient, reproducible siRNA transfections in different cell types.

Furthermore, both can be used to transfect cells during subculturing-saving a full day of valuable time-without increased cytotoxicity. In addition to containing a choice of transfection agents, this kit includes a well-characterized GAPDH synthetic siRNA that can be used to optimize transfection conditions. Results can also be verified at the protein level with GAPDH-specific antibodies. A scrambled, negative control siRNA is also provided to confirm the specificity of the siRNA of interest and to demonstrate the absence of nonspecific, toxic effects.

For Research Use Only. Not for use in diagnostic procedures.
Specifications
Control TypeNegative Control, Positive Control
Cell TypeMammalian
FormatKit
Sample TypeSynthetic siRNA, miRNA
Transfection TechniqueLipid-based Transfection
For Use With (Application)siRNA Transfection
Product LineSilencer™, Ambion™
Product TypesiRNA Transfection Optimization Kit
Quantity1 mL
RNAi TypesiRNA
Unit SizeEach
Contents & Storage
siPORT™ NeoFX™ Transfection Agent and siPORT™ Amine Transfection Agent should be stored at 4°C. GAPDH siRNA and Negative Control siRNA should be stored at -20°C.

Frequently asked questions (FAQs)

What are the benefits of using a vector to deliver RNAi?

Vector technologies allow you to:

Achieve transient or stable target knockdown
Perform RNAi in any cell type, even hard-to-transfect, primary, and non-dividing cells
Regulate gene inhibition with inducible siRNA expression
Select for a pure population of cells stably expressing an siRNA sequence
Control gene expression in vivo with tissue-specific promoters

Find additional tips, troubleshooting help, and resources within our RNAi Support Center.

Why are my cells dying after transfection?

We would suggest running a transfection reagent control only to determine if your cells are sensitive to the transfection reagent. Additionally, you can try using different cell densities and siRNA concentrations to diminish any toxic effects from the transfection itself.

Find additional tips, troubleshooting help, and resources within our RNAi Support Center.

I transfected my siRNA and the mRNA levels are down, but the protein is not. Why is that?

In some cases, knockdown of a protein can be affected by other variables such as protein turnover rate, even though the RNA is knocked down. Additionally, a longer time course may be needed to see an effect on protein compared to mRNA.

Find additional tips, troubleshooting help, and resources within our RNAi Support Center.

I am not getting my target knockdown. What could be the cause of this?

Please see the following possibilities and suggestions:

- How many siRNA did you test? Is there any knockdown? If there is no knockdown (<10%) in any of the siRNA, then the assay is likely the problem. Try using a different qRT-PCR assay to assess knockdown.
- What was the positioning of the qRT-PCR assay target site relative to the cut site for the siRNA? If greater than 3,000 bases away, the problem could be alternative splice transcripts.
- What are the Cts for the experiment? They should be below 35 in a 40-cycle qRT-PCR experiment.
- Did you confirm the siRNA got into the cell? We recommend using a validated positive control siRNA to check the transfection efficiency.

Find additional tips, troubleshooting help, and resources within our RNAi Support Center.

I am not getting any knockdown with my siRNA. What do you suggest I try?

Please see the following possibilities and suggestions:

- Were the mRNA levels checked? The most reliable method is real-time PCR. In some cases, knockdown of a protein can be affected by other variables, such as protein turnover rate, even though the RNA is knocked down.
- How is the RNA being isolated? Has the quality of the isolated RNA been checked? Ensure that the RNA has not been degraded.
- Was a positive control used? This can help to determine whether the reagents are working and whether the siRNA was delivered correctly to the cell. Run your experiment in parallel with the positive control siRNA.
- Was a transfection control used? What is the percentage of transfected cells?
- Was a time course used? Generally, gene silencing can be assessed as early as 24 hours posttransfection. However, the duration and level of knockdown are dependent on cell type and concentration of siRNA.
- Was optimization of transfection conditions performed? You can try using different cell densities and siRNA concentrations.
- Which concentration of siRNA did you use? We recommend testing multiple concentrations between 5 nM and 100 nM.

Find additional tips, troubleshooting help, and resources within our RNAi Support Center.

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