Stealth RNAi™ siRNA Negative Control Kit

Citas y referencias (3)

Invitrogen™

Stealth RNAi™ siRNA Negative Control Kit

Los controles negativos ARNip Stealth RNAi™ constituyen un método excelente para medir el efecto de su dúplex de ARNip experimentalMás información

Número de catálogo	Cantidad
12935100	1 Kit

Número de catálogo 12935100

Precio (MXN)

Cantidad:

1 Kit

Los controles negativos ARNip Stealth RNAi™ constituyen un método excelente para medir el efecto de su dúplex de ARNip experimental Stealth RNAi™ con respecto al fondo. Estos controles:

• Están diseñados con tres niveles de contenido de GC (bajo, medio y alto) y con tres secuencias disponibles para cada nivel, de modo que los investigadores pueden igualar el contenido de GC con su ARNip experimental Stealth RNAi™
• No son homólogos a ningún elemento del transcriptoma de vertebrados
• Están probados para no inducir respuesta de estrés ARNip Stealth RNAi™

El kit de control negativo contiene los tres controles (de bajo, medio y alto contenido de GC, excluyendo las secuencias n.° 2 y 3) y cada uno también está disponible por separado. Obtenga los controles negativos ideales para su experimento de RNAi para que pueda evaluar sus resultados con precisión.

Para uso exclusivo en investigación. No apto para uso en procedimientos diagnósticos.

Especificaciones

Para utilizar con (aplicación)Transfección, ARNi

Etiqueta o tinteno conjugado

Línea de productosARNip Stealth RNAi

Tipo de productoARNip

Cantidad1 Kit

Tipo de controlControl negativo

RNAi TypeARNip

Unit SizeEach

Contenido y almacenamiento

Cada control individual se suministra como 250 µl a 20 µM. El kit de control negativo de ARNip Stealth RNAi™ contiene uno de cada uno de los tres controles negativos (cada uno 250 µl a 20 µM). Almacenar a -20 °C. Se garantiza la estabilidad de todos los productos durante 6 meses si se almacenan adecuadamente.

Preguntas frecuentes

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.

View all Stealth RNAi™ siRNA Negative Control Kit FAQs

Citations & References (3)

Citations & References

Abstract

The Heat Shock Protein 90-CDC37 Chaperone Complex Is Required for Signaling by Types I and II Interferons.

Authors:Shang L, Tomasi TB,

Journal:J Biol Chem

PubMed ID:16280321

Interferon signaling pathways are critical to both innate and adaptive mmunity. We have demonstrated here that the inhibition of heat shock protein 90 (Hsp90) functions by small interfering RNAs or chemical inhibitors blocking interferon-induced gene expression. Hsp90 was required for signal transducers and activators of transcription 1 phosphorylation, and in ... More

NOD2/CARD15 mediates induction of the antimicrobial peptide human beta-defensin-2.

Authors:Voss E, Wehkamp J, Wehkamp K, Stange EF, Schröder JM, Harder J,

Journal:J Biol Chem

PubMed ID:16319062

Production of inducible antimicrobial peptides offers a first and rapid defense response of epithelial cells against invading microbes. Human Beta-defensin-2 (hBD-2) is an antimicrobial peptide induced in various epithelia upon extracellular as well as intracellular bacterial challenge. Nucleotide-binding oligomerization domain protein 2 (NOD2/CARD15) is a cytosolic protein involved in intracellular ... More

Molecular cloning and characterization of a novel 3'-phosphoadenosine 5'-phosphosulfate transporter, PAPST2.

Authors:Kamiyama S, Sasaki N, Goda E, Ui-Tei K, Saigo K, Narimatsu H, Jigami Y, Kannagi R, Irimura T, Nishihara S,

Journal:J Biol Chem

PubMed ID:16492677

Sulfation is an important posttranslational modification associated with a variety of molecules. It requires the involvement of the high energy form of the universal sulfate donor, 3'-phosphoadenosine 5'-phosphosulfate (PAPS). Recently, we identified a PAPS transporter gene in both humans and Drosophila. Although human colonic epithelial tissues express many sulfated glycoconjugates, ... More