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Ambion™ RNase Inhibitor, cloned, 40 U/µL (Invitrogen™)

Ambion® RNase Inhibitor, a recombinant human protein produced in E. coli, is a potent inhibitor of neutral pancreatic RNase A type enzymes. The mode of inhibition is noncompetitive; the inhibitor tightly binds RNases in a 1:1 ratio. Supplied in one tube containing 2,500 U (40 U/ µL). The enzyme has been shown to inactivate RNases present in many tissues and cell types. Addition of the ribonuclease inhibitor (RI) has been shown to be useful whenever the integrity of RNA must be maintained such as in the preparation of cDNA by reverse transcription, in vitro RNA transcription, and in in vitro protein synthesis. RI does not inhibit RNase I, T1, T2, H, U1, U2, or CL3. RI requires a minimum of 1 mM DTT to maintain activity, and has an activity range between pH 5.0 to 8.0 with maximal activity between pH 7.0 and 8.0. Since the mode of inhibition is the formation of a 1:1 complex with RNases, care must be taken to avoid denaturation or oxidation of the ribonuclease inhibitor (e.g. by addition of SDS, urea, etc.). RI should be added to transcription, translation, and cDNA synthesis reactions to give a final concentration of 1 U/ µL. RNase Inhibitor is rigorously tested for contaminating RNase, exonuclease, endonuclease, and protease activity.

Unit Definition:
One unit is the amount of protein required to inhibit the activity of 5 ng of RNase A by 50%. Unit assay conditions: 100 mM Tris-acetate (pH 6.5), 1 mM EDTA, 1 mM cyclic 2',3'-CMP, and RNase inhibitor. Addition of RNase A initiates the reaction. RNase Inhibitor activity is measured by the inhibition of hydrolysis of cyclic 2', 3'-CMP by RNase A.

Ambion™ In Vivo Negative Control #1 siRNA (Invitrogen™)

Ambion® In Vivo siRNA Controls
• Validated siRNA controls for optimizing siRNA experiments targeting coding and noncoding transcripts
• Positive Control siRNAs functionally tested in several common cell lines
• Negative Controls functionally proven to have minimal effects on cell proliferation and viability
• Include Silencer® Select modifications for enhanced specificity or Ambion® In Vivo modifications for increased stability
• For use in human, mouse, and rat cells

What are Ambion® In Vivo siRNAs?
Ambion® In Vivo siRNAs are designed using the proven Silencer® Select algorithm and incorporate chemical modifications for superior serum stability with in vivo delivery. The added serum stability does not compromise the performance of the siRNAs. Ambion® In Vivo siRNAs have been shown to be non-toxic in vivo (mouse) and non-immunogenic (cell-based assays). In cell-based assays, Ambion® In Vivo siRNAs exhibit equivalent or better potency than Silencer® Select siRNAs. The result is better confidence in your in vivo RNAi experiments, better knockdown when paired with an efficient delivery solution, and the assurance of limited non-specific effects.

Negative Control siRNAs
Negative control siRNAs—siRNAs with sequences that do not target any gene product—are essential for determining the effects of siRNA delivery and for providing a baseline to compare siRNA-treated samples. Ambion® In Vivo siRNAs and have no significant sequence similarity to mouse, rat, or human gene sequences. These negative control siRNAs have been tested by microarray analysis and shown to have minimal effects on gene expression. In addition, these negative control siRNA has been tested in multi-parametric cell-based assays and are proven to have no significant effect on cell proliferation, viability, or morphology in the cell lines tested.

Quality Control
Ambion® synthesizes and purifies each Ambion® In Vivo siRNA in state-of-the-art facilities to meet the highest quality standards. As part of our rigorous quality control procedures, each RNA oligonucleotide is analyzed by MALDI-TOF mass spectrometry, and analytical HPLC is used to monitor purity. To provide the utmost in quality, we also assess each annealed siRNA by gel electrophoresis to confirm that the strands anneal properly. The result is premium-quality siRNA that is purified and ready to use.

In Vivo Ready Controls
Our "In Vivo Ready" Control siRNAs are subjected to that extra level of purification and testing required for the introduction of siRNAs into animals. After HPLC purification and annealing, each siRNA is further purified utilizing a process that removes excess salt via a semi-permeable 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 presence of deionized water, in vivo Ready siRNAs contain <5.0 mM Na+, <0.06 mM K+, and <0.02 mM Mg2+.

Ambion™ In Vivo GAPDH Positive Control siRNA (Invitrogen™)

Ambion® In Vivo siRNA Controls
• Validated siRNA controls for optimizing siRNA experiments targeting coding and noncoding transcripts
• Positive Control siRNAs functionally tested in several common cell lines
• Negative Controls functionally proven to have minimal effects on cell proliferation and viability
• Include Silencer® Select modifications for enhanced specificity or Ambion® In Vivo modifications for increased stability
• For use in human, mouse, and rat cells

What are Ambion® In Vivo siRNAs?
Ambion® In Vivo siRNAs are designed using the proven Silencer® Select algorithm and incorporate chemical modifications for superior serum stability with in vivo delivery. The added serum stability does not compromise the performance of the siRNAs. Ambion® In Vivo siRNAs have been shown to be non-toxic in vivo (mouse) and non-immunogenic (cell-based assays). In cell-based assays, Ambion® In Vivo siRNAs exhibit equivalent or better potency than Silencer® Select siRNAs. The result is better confidence in your in vivo RNAi experiments, better knockdown when paired with an efficient delivery solution, and the assurance of limited non-specific effects.

GAPDH Positive Control siRNAs
Our extensively validated, positive control siRNA to human, mouse, and rat GAPDH serves multiple functions. First, it is an ideal “test" siRNA for those just beginning siRNA experiments, because it is validated to work in multiple cell lines. In addition, because it targets GAPDH mRNA, which is commonly used as an internal control, its effects are easy to assay, and thus provides an excellent tool to monitor siRNA transfection efficiency by real-time RT-PCR. The Ambion® In Vivo GAPDH Positive Control siRNA shows increased serum and nuclease resistance compared with Silencer® Select siRNAs without loss of potency or specificity and is recommended for in vivo applications.

Quality Control
Ambion® synthesizes and purifies each Ambion® In Vivo siRNA in state-of-the-art facilities to meet the highest quality standards. As part of our rigorous quality control procedures, each RNA oligonucleotide is analyzed by MALDI-TOF mass spectrometry, and analytical HPLC is used to monitor purity. To provide the utmost in quality, we also assess each annealed siRNA by gel electrophoresis to confirm that the strands anneal properly. The result is premium-quality siRNA that is purified and ready to use.

In Vivo Ready Controls
Our "In Vivo Ready" Control siRNAs are subjected to that extra level of purification and testing required for the introduction of siRNAs into animals. After HPLC purification and annealing, each siRNA is further purified utilizing a process that removes excess salt via a semi-permeable 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 presence of deionized water, in vivo Ready siRNAs contain <5.0 mM Na+, <0.06 mM K+, and <0.02 mM Mg2+.

Ambion™ Recombinant RNase A (Invitrogen™)

Ambion® Recombinant RNase A for the production of plasmid DNA that is substantially free of host RNA is now available for research applications. For large batch sizes, the use of RNase is still the only effective technique applicable to remove host RNA. Other methods such as selective precipitation, gel filtration, chromatography, and endogenous E. coli RNase digestion have only been effective for small-scale or bench-scale purification. This new product was designed specifically to minimize the risk of introducing pathogens into bioprocessing. Recombinant RNase A is rigorously tested for contaminating nonspecific endonuclease, exonuclease, and protease activity.

Ambion™ T4 RNA Ligase, cloned, 5 U/µL (Invitrogen™)

Ambion® T4 RNA Ligase catalyzes the formation of a phosphodiester linkage between a 5'-phosphoryl-terminated nucleic acid donor and a 3'-hydroxyl-terminated nucleic acid acceptor. Substrates include RNA, DNA, and oligonucleotides. One tube containing 2,500 U (5 U/ µL) is provided along with 10X Reaction Buffer. The enzyme can be used for tagging the 5' ends of mRNA with oligonucleotides for mapping studies (RACE), 3'-end labeling RNA molecules, and circularizing RNA and DNA molecules.

Unit Definition:
One unit catalyzes the formation of 1 nmol of [5'-32P]-rA12-18 into a phosphatase-resistant form in 30 min at 37°C.

Ambion™ In Vivo Factor VII Positive Control siRNA (Invitrogen™)

Ambion® In Vivo siRNA Controls
• Validated siRNA controls for optimizing siRNA experiments targeting coding and noncoding transcripts
• Positive Control siRNAs functionally tested in several common cell lines
• Negative Controls functionally proven to have minimal effects on cell proliferation and viability
• Include Silencer® Select modifications for enhanced specificity or Ambion® In Vivo modifications for increased stability
• For use in mouse cells

What are Ambion® In Vivo siRNAs?
Ambion® In Vivo siRNAs are designed using the proven Silencer® Select algorithm and incorporate chemical modifications for superior serum stability with in vivo delivery. The added serum stability does not compromise the performance of the siRNAs. Ambion® In Vivo siRNAs have been shown to be non-toxic in vivo (mouse) and non-immunogenic (cell-based assays). In cell-based assays, Ambion® In Vivo siRNAs exhibit equivalent or better potency than Silencer® Select siRNAs. The result is better confidence in your in vivo RNAi experiments, better knockdown when paired with an efficient delivery solution, and the assurance of limited non-specific effects.

Factor VII (FVII) Positive Control siRNA
Ambion® In Vivo Factor VII siRNA, In Vivo Ready, provides a positive control for experiments involving Ambion® In Vivo siRNA delivery to mice. Factor VII (FVII⁄F7; also known as proconvertin) is a vitamin K-dependent serine protease that functions as a central protein in the coagulation cascade. FVII is synthesized in the liver and secreted to the plasma where it circulates in an inactive form. After delivery of FVII-targeting siRNA to the liver, knockdown can be measured by quantifying FVII protein levels in blood by robust, commercially available chromogenic assays. Moreover, blood samples can be collected from the same animal at multiple time points post-siRNA injection. These features enable much more rapid and efficient knockdown measurements for FVII than for gene targets that require quantitation of mRNA levels in liver tissue, making FVII an ideal control for in vivo siRNA delivery.

Quality Control
Ambion® synthesizes and purifies each Ambion® In Vivo siRNA in state-of-the-art facilities to meet the highest quality standards. As part of our rigorous quality control procedures, each RNA oligonucleotide is analyzed by MALDI-TOF mass spectrometry, and analytical HPLC is used to monitor purity. To provide the utmost in quality, we also assess each annealed siRNA by gel electrophoresis to confirm that the strands anneal properly. The result is premium-quality siRNA that is purified and ready to use.

In Vivo Ready Controls
Our "In Vivo Ready" Control siRNAs are subjected to that extra level of purification and testing required for the introduction of siRNAs into animals. After HPLC purification and annealing, each siRNA is further purified utilizing a process that removes excess salt via a semi-permeable 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 presence of deionized water, in vivo Ready siRNAs contain <5.0 mM Na+, <0.06 mM K+, and <0.02 mM Mg2+.

Ambion™ RNase A, affinity purified, 1 mg/mL (Invitrogen™)

Ambion® RNase A is an endonuclease that specifically cleaves 3' of U and C residues. Affinity Purified RNase A is intended for critical applications when the absence of DNase and other nonspecific nuclease activities is essential. Supplied in one tube containing 200 µg (1 mg/mL). Note: This preparation contains RNase B, a carbohydrate isoform of RNase A. RNase A is rigorously tested for contaminating nonspecific endonuclease, exonuclease, and protease activity.

Ambion™ DNase I (RNase-free) (Invitrogen™)

Ambion DNase I (RNase-free) (E.C. 3.1.21.1) is a nonspecific endonuclease that degrades double- and single-stranded DNA and chromatin. It functions by hydrolyzing phosphodiester linkages, producing mono and oligonucleotides with a 5'-phosphate and a 3'-hydroxyl group. RNase-free DNase I is of the highest purity available and is recommended to degrade DNA in the presence of RNA when the absence of RNase is critical to maintain the integrity of the RNA. For example, DNase I is frequently used to remove template DNA following in vitro transcription, and to remove contaminating DNA in total RNA preparations (especially those from transfected cells that may contain plasmid DNA), used for ribonuclease protection assays, cDNA library contraction, and RT-PCR. DNase I requires bivalent cations (Mg2+ and Ca2+ at approximately 5 mM and 0.5 mM, respectively) for maximal activity, and has a pH optimum of 7.8.

RNase-free DNase I outperforms the competition
A research report in BioTechniques (Matthews et al., 32: 1412-1417, 2002) compared RNase contamination in DNase I preparations from Sigma, Roche, Applied Science, Qiagen, and Ambion. The results revealed that "...with the exception of Ambion®'s RNase-free DNase I, the integrity of cRNA from in vitro transcription reactions was compromised and was still contaminated with DNA. Ambion®'s DNase was used for the remaining experiments requiring DNase digestion...". Ambion® DNase I is tested for contaminating RNase and protease activity. Functionality is determined by digestion of human genomic DNA followed by quantitative real-time PCR to detect undigested DNA.

Unit definition
One unit is the amount of enzyme required to completely degrade 1 µg DNA in 10 min at 37°C, and is equivalent to 0.04 Kunitz units.

Accessory products
For an alternative to bovine DNase I, please consider Recombinant DNase I (Cat. No. AM2235). For a more-active, salt-tolerant DNase, please see the TURBO™ DNase products (Cat. Nos. AM2239 and AM2238).

Ambion™ RNase I, cloned, 100 U/µL (Invitrogen™)

Ambion® RNase I efficiently cleaves after all four bases of single-stranded RNA, in contrast to RNase A, which only cleaves after C and U residues. RNase I degrades all RNA dinucleotide bonds leaving a 5' hydroxyl and 2', 3' cyclic monophosphate. Supplied in one tube of 10,000 U (100 U/ µL). RNase I will degrade any RNA to a mixture of mononucleotides, dinucleotides, and trinucleotides and does not degrade DNA, although it will bind to DNA. It has a marked preference for single-stranded RNA over double-stranded RNA, which allows it to work well in ribonuclease protection assays. This RNase I is rigorously tested for nonspecific contaminating endonuclease, exonuclease, and protease activity.

Unit Definition:
One unit is the amount of enzyme required to produce 1 µg of acid-soluble material from mouse liver RNA in 30 min at 37°C.

Ambion™ RNase III (Invitrogen™)

Ambion® Escherichia coli ribonuclease III (RNase III; EC 3.1.24) is a double-stranded RNA (dsRNA) specific endoribonuclease. In E. coli, RNase III cleaves ribosomal RNA (rRNA) precursors during maturation of rRNA. The enzyme cleaves dsRNA into 12–15 bp dsRNA fragments with 2 to 3 nucleotide 3' overhangs, and 5' phosphate and 3' hydroxyl termini. The termini and overhangs of RNase III cleavage products are thus the same as those produced by Dicer in the eukaryotic RNAi pathway. Transfection of RNase III cleavage products can be used to induce RNAi in mammalian cells (patent pending).

Unit Definition:
One unit is defined as the amount of enzyme catalyzing the cleavage of 1 µg of 500 bp dsRNA substrate to approximately 12–30 bp fragments in 60 min at 37°C.

Custom Designed Ambion™ In Vivo (Invitrogen™)

RNA interference (RNAi) is the best way to effectively knock down gene expression to study protein function in a wide range of cell types. Use our GeneAssist™ Custom siRNA Builder to design and order a custom siRNA.

Ambion™ In Vivo Pre-Designed siRNA (Invitrogen™)

RNA interference (RNAi) is the best way to effectively knock down gene expression to study protein function in a wide range of cell types. Use our GeneAssist™ Custom siRNA Builder to design and order a custom siRNA.

Ambion™ T7 RNA Polymerase, cloned, 200 U/µL (Invitrogen™)

This cloned high-purity Ambion® T7 RNA Polymerase has a high specificity for its promoter. The enzyme will transcribe large amounts of RNA from DNA sequences downstream of the promoter and show no cross talk between promoters. One tube containing 30,000 U at a concentration of 200 U/ µL is provided. Its high purity makes them ideal for synthesizing high specific activity RNA hybridization probes, biologically active mRNA, and antisense RNA. The RNA Polymerase is rigorously tested for contaminating nonspecific endonuclease, exonuclease, protease, and RNase activity. It is also tested for functionality with the MAXIscript® Kit at 20 U per reaction.

Unit Definition:
One unit of RNA Polymerase will catalyze the incorporation of 1 nmol of nucleoside triphosphate into acid-insoluble material in 60 min at 37°C.

Ambion™ RNase H, from E. coli, 10 U/µL (Invitrogen™)

Ambion® RNase H (Ribonuclease H) is isolated from an E. coli strain that over-expresses the gene. RNase H specifically degrades the RNA in RNA:DNA hybrids to produce 3' -hydroxyl and 5' -phosphate terminated products. It is supplied in one tube containing 1,000 U (10 U/ µL). The enzyme will not degrade DNA or unhybridized RNA. RNase H is an integral part of most RNA amplification and NASBA protocols. It can also be used to degrade specific RNAs when the complementary DNA oligo is hybridized, such as poly(A) tail removal from mRNA hybridized to oligo(dT). Ribonuclease H is rigorously tested for contaminating nonspecific endonuclease, exonuclease, RNase, and protease activity.

Unit Definition:
One unit of Ribonuclease H is the amount of enzyme required to increase fluorescence 1.5 RFUs per sec at 37°C using 20 pmol of RNaseAlert® probe coupled to 1,000 pmol of a complementary oligonucleotide as substrate.

Ambion™ In Vivo Pre-Designed siRNA (Invitrogen™)

Ambion® siRNAs are classic 21-mers which incorporate the latest improvements in siRNA design, off-target effect prediction algorithms, and chemistry. These siRNAs are guaranteed-to-silence based on their proven design.