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Invitrogen™ Silencer™ GAPDH Positive Control siRNA, in vivo ready

Catalog No. 4404025
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4404025 250 nmol
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Catalog No. 4404025 Supplier Invitrogen™ Supplier No. 4404025
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Description

Ambion™ Silencer™ GAPDH Positive Control siRNA, In Vivo Ready is ideal for developing and optimizing siRNA experiments.

Ambion™ Silencer™ GAPDH Positive Control siRNA, In Vivo Ready is ideal for developing and optimizing siRNA experiments. The control is validated for use in human cell lines and optimized for mouse and rat. It is highly purified as required for animal use. 250 nmol is provided.

  • Validated siRNA controls for optimizing siRNA experiments
  • Gene-specific control siRNAs provided with scrambled, negative controls
  • Functionally tested in several common cell lines
  • HPLC purified, duplexed, and ready to use

Quality Control:

Silencer™ siRNAs are manufactured in a state-of-the-art facility under the highest quality standards. After HPLC purification and annealing, each siRNA is further purified utilizing a process that removes excess salt via a semipermeable membrane. The result is highly pure siRNA with minimal salt content, suitable for in vivo applications. In vivo siRNAs are then sterile filtered and tested for the presence of endotoxin. At concentrations of 50 μM in deionized water, In Vivo Ready siRNAs contain less than 0.6 mM Na+, less than 2.0 mM K+, and less than 0.1 mM Mg2+. Silencer™ control siRNAs are unmodified 21-mers; they are designed to be used with Silencer™ siRNAs, BLOCK-iT™ siRNAs, and other unmodified 21mer siRNAs. If you are using other siRNA products, such as Ambion™ Silencer™ Select siRNAs or STEALTH RNAi™ siRNAs, we recommend that you use control siRNAs designed for use with those siRNAs for more reliable experimental results.

Specifications

Content And Storage The siRNA is provided dried down in a single tube, which should be stored at -20°C.
Description The siRNA is provided dried down in a single tube, along with Nuclease-free Water for resuspension, and should be stored at –20°C.
Control Type Gene Knockdown Positive Control
RNAi Type siRNA
For Use With (Application) RNAi
Label or Dye Unconjugated
Product Line Silencer, Ambion
Product Type siRNA
Purity HPLC
Quantity 250 nmol
Shipping Condition Room Temperature
Target GAPDH
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Frequently Asked Questions (FAQs)

What controls should I use for my in vivo siRNA studies?

We offer the following in vivo siRNA controls:

Invitrogen In Vivo Negative Control #1 siRNA (Cat. No. 4457287, 5nmol; 4459405, 50 nmol; 4457289, 250 nmol)
Invitrogen In Vivo GAPDH Positive control siRNA (Cat. No. 4457288, 5 nmol; 4459407, 50 nmol, 4457291, 250 nmol)
Invitrogen In Vivo Factor VII Positive control siRNA (Cat. No. 4459408, 50 nmol; 4457292, 250 nmol)
Silencer Select Negative Control No. 1 siRNA, in vivo ready (Cat. No. 4404020, 250 nmol)
Silencer GAPDH Positive Control siRNA, in vivo ready (Cat. No. 4404025, 250 nmol)

How stable is Stealth siRNA, can I use it for in vivo research?

Stealth siRNA is markedly more stable in serum than unmodified siRNA. This increased stability is particularly beneficial in vivo where the RNAi may be exposed to more proteases. In collaboration with Intradigm, Stealth RNAi has been demonstrated to have activity in vivo when injected intra-tumorally.
How can I order in vivo siRNA duplexes?

Pre-designed Stealth RNAi and Ambion In Vivo siRNA can be ordered directly from our pre-designed siRNA search interface (https://www.thermofisher.com/us/en/home/life-science/rnai/introduction-to-in-vivo-rnai.html?open=faq#search) to target human, mouse, or rat genes.

Custom siRNA designs for in vivo experiments can be ordered from two interfaces, depending on the product format being requested: Custom Ambion In Vivo siRNA is ordered from the GeneAssist Custom siRNA Builder (https://www.thermofisher.com/order/custom-genomic-products/tools/sirna/), while Stealth RNAi is ordered using the BLOCK-iT RNAi Express for in vivo synthetics (https://rnaidesigner.thermofisher.com/rnaiexpress/inVivoService.jsp).
Do I need endotoxin testing for my in vivo RNAi duplexes?

It is recommended to reduce the chance of eliciting an immune response.
I want to order larger scales than the standard offering, whom do I contact?

Please contact rnairesearcher@invitrogen.com regarding large scale in vivo siRNA or fill out the quote request form (www.thermofisher.com/us/en/home/global/forms/custom-sirna-mirna-quote-request.html).

How should I deliver my in vivo RNAi molecules?

Several different approaches have been used for siRNA delivery, including various local delivery techniques and systemic delivery. We offer a lipid reagent, Invivofectamine 3.0 Transfection Reagent for in vivo siRNA systemic delivery. Learn more about this reagent here (https://www.thermofisher.com/us/en/home/life-science/rnai/introduction-to-in-vivo-rnai/invivofectamine-reagent.html).
Should I use siRNA or vectors for in vivo RNAi?

RNAi can be delivered using two different approaches-siRNA synthetic duplexes or siRNA expressed from plasmids or viral vectors (shRNA, miRNAi). siRNA are becoming the method of choice for the fast development of therapeutics. They are easy to use, easy to design, and easy to synthesize. siRNA can be rapidly identified and multiple genes can be targeted at the same time. With RNAi vectors, the expression will be steadier as a result of the possibility of stable integration of the plasmid into the genome, and they have the ability to target nondividing cells such as stem cells, lymphocytes and neurons. The drawbacks are the danger of oncogenic transformation from insertional mutagenesis, and unanticipated toxicity from long-term silencing of human genes and/or having high amounts of siRNA inside the cell (Grimm D. et al.: Nature 441: 537-541 (2006)).

I am planning on using siRNA for my in vivo experiments, what purity should they be?

For Stealth siRNA we recommend to select HPLC in vivo purity. For Invitrogen In Vivo siRNA, we recommend in vivo ready purity.
Should I use labeled or unlabeled siRNA for my in vivo siRNA experiments?

We have demonstrated that labeling Stealth and Silencer siRNA does not hamper their knockdown potency. An alternative approach is to mix unlabeled duplexes with labeled control duplexes; this method is more commonly used with in vivo siRNA, and allows progression to clinical research unhindered by questions about the possible effects of a label.

Should I use chemically modified duplexes for my in vivo siRNA experiments?

Chemically modified siRNA duplexes have a number of advantages over standard siRNA duplexes, including the minimization of off-target effects, enhanced stability, and reduced toxicity. For these reasons, chemically modified siRNA duplexes are recommended for in vivo siRNA experiments. For some pilot experiments, where the only aim is to determine biodistribution, unmodified siRNA and fluorescent controls are useful and slightly more cost-effective.
What are the benefits of using our Invitrogen In vivo siRNA?

- 100x more stable than unmodified siRNA - longer knockdown in vivo
- Compatible with any 21-mer sequence - no loss of potency
- Non-toxic - actually reduces siRNA sequence specific immune responses
- Enhanced specificity - up to 90% reduction in off target effects
- Both Pre-designed and Validated Invitrogen In Vivo siRNAs offered
What siRNA do you offer that I can use for my in vivo studies, and what are the differences between them?

Please see the different siRNA we offer for in vivo studies:

Ambion In Vivo Pre-designed siRNAs are designed using the Silencer Select algorithm and incorporate additional chemical modifications for superior serum stability (half life >5 hours at 37 degrees C in 90% mouse serum) with in vivo applications. Ambion In Vivo siRNAs are non-toxic and non-immunogenic in vitro (peripheral blood nononuclear cells; PBMC) and in vivo (mouse). In cell-based assays, Ambion In Vivo siRNAs exhibit potency and specificity equivalent to Silencer Select siRNAs.
Ambion In Vivo Custom Designed siRNAs are designed to your specified target, using the Silencer Select algorithm and Ambion In Vivo chemical modifications.
Ambion In Vivo Custom Synthesized siRNAs are synthesized with your sequence, with Ambion In Vivo chemical modifications for serum stability.

Our Ambion In Vivo siRNA can be ordered with the following purities:

Desalted: Desalted and analyzed by MALDI-TOF mass spectrometry.
HPLC: HPLC purified and analyzed by MALDI-TOF and analytical HPLC
In Vivo Ready : HPLC purified, analyzed by MALDI-TOF and analytical HPLC, dialyzed to remove salts, sterile filtered, and endotoxin tested

Use our GeneAssist tool to order your custom siRNA (http://www.thermofisher.com/order/custom-genomic-products/tools/sirna/).

How critical is it to include positive and negative controls in siRNA experiments?

A positive control siRNA such as the Silencer Select GAPDH siRNA will demonstrate efficient transfection and help to optimize transfection conditions. A negative control siRNA is necessary to serve as a baseline for gene expression.
What are the positive and negative controls I should use with my Silencer or Silencer Select siRNA experiment?

We offer several negative and positive controls for both Silencer and Silencer Select siRNA.

For Silencer siRNA positive control:
- Silencer Firefly Luciferase (GL2 + GL3) siRNA (Cat. No. AM4629, 5nmol)
- Silencer GAPDH siRNA (human) (Cat. No AM4605, 5nmol; AM4633, 40 nmol)
- Silencer GAPDH siRNA (human, mouse, rat) (Cat. No. AM4624, 5 nmol; AM4631, 40 nmol; AM4632, 5 x 40 nmol)
- Silencer GFP (eGFP) siRNA (Cat. No. AM4626, 5 nmol)
- Silencer KIF11 (Eg5) siRNA (human, mouse, rat) (Cat. No. AM4639, 5 nmol)

If working with Fluorescently labeled Silencer siRNA:
- Silencer Cy-3 labeled GAPDH siRNA (human, mouse, rat) (Cat. No. AM4649, 5 nmol)
- Silencer FAM-labeled GAPDH siRNA (human, mouse, rat) (Cat. NO. AM4650, 5 nmol)

If working with Silencer siRNA in vivo ready:
- Silencer GAPDH Positive Control siRNA, in vivo ready (Cat. No. 4404025, 250 nmol)

For Silencer siRNA negative controls:
- Silencer Negative Control No. 1 siRNA (Cat. Nos. AM4611, 5 nmol; AM4635, 40 nmol; AM4636, 5 x 40 nmol)
- Silencer Negative Control No. 2 siRNA (Cat. Nos. AM4613, 5 nmol; AM4637, 30 nmol)
- Silencer Negative Control No. 3 siRNA (Cat. No. AM4615, 5 nmol)
- Silencer Negative Control No. 4 siRNA (Cat. No. AM4641, 5 nmol)
- Silencer Negative Control No. 5 siRNA (Cat. No. AM4642, 5 nmol)

If working with Fluorescently labeled Silencer siRNA:
- Silencer Cy 3-labeled Negative Control No. 1 siRNA (Cat. No AM4621, 5 nmol)
- Silencer FAM-labeled Negative Control No. 1 siRNA (Cat. No. AM4620, 5 nmol)

If working with Silencer siRNA in vivo ready:
- Silencer in vivo ready (Cat. No. 4404021)

For Silencer Select siRNA Positive control:
- Silencer Select GAPDH Positive Control siRNA (Cat. No. 4390849, 5 nmol; 4390850, 40 nmol; 4404024, 250 nmol)
- Silencer Select MALAT1 Positive Control siRNA for non-coding siRNA (Cat. No. 4455877, 5 nmol)

For Silencer Select siRNA negative control:
- Silencer Select Negative Control No. 1 siRNA (Cat. No. 4390843, 5 nmol; 4390844, 40 nmol)
- Silencer Select Negative Control No. 2 siRNA (Cat. No. 4390846, 5 nmol; 4390847, 40 nmol)

If working with Silencer Select siRNA in vivo ready:
- Silencer Select Negative Control No. 1 siRNA, in vivo ready (Cat. No. 4404020, 250nmol)

For Research Use Only. Not for use in diagnostic procedures.