2012 Promega catalogue

Cell Signaling
RNasin ® Plus RNase Inhibitor
Product Size Conc. Cat.# Price ($)
RNasin ® Plus RNase Inhibitor 2,500 u 40 u/µl N2611 173.00
For Laboratory Use.
10,000 u 40 u/µl N2615 597.00
Description: RNasin ® Plus RNase Inhibitor is a recombinant mammalian
RNase inhibitor that is expressed as a soluble protein in E. coli, allowing easy
purification through a combination of ion exchange and hydrophobic interaction
chromatography. The protein is capable of inhibiting eukaryotic RNases (e.g.,
RNase A and RNase B) similarly to human placental RNase inhibitor. RNasin ®
Plus RNase Inhibitor is tested in RT-PCR and is compatible with enzymes such
as AMV, M-MLV and ImProm-II Reverse Transcriptases or Taq and Tfl DNA
Polymerases. RNasin ® Plus RNase Inhibitor also is tested and compatible with
quantitative, real-time RT-PCR in a TaqMan ® assay.
The inhibitor offers increased resistance to oxidation over the human version of
the protein. Two cysteines in the human protein have been identified as
especially sensitive to oxidation and react by forming a disulfide bond that can
block the active site of the inhibitor. RNasin ® Plus, through natural amino acid
diversity, lacks the ability to form this site-blocking disulfide. In addition, the
new protein has characteristics never before realized, including continued
inhibition of RNases above 50°C. Heating solutions of RNasin ® Plus and RNase
followed by cooling does not result in the reappearance of RNase activity—
even when the solution is heated above the denaturation temperature of the
RNasin ® Plus protein alone. This allows RNasin ® Plus to protect RNA species
prior to, during and after heating, even at temperatures normally used during
first-strand DNA synthesis in RT-PCR. We have taken solutions up to 70°C for
15 minutes and did not see RNase reactivation.
Features:
• Improved Resistance to Oxidation: Due to natural amino acid diversity,
RNasin ® Plus lacks the capability to form the active site-blocking disulfide
bond that can form in the human protein under oxidative conditions.
• Improved Purification: RNasin ® Plus is expressed by E. coli as a soluble
protein, allowing easy purification by a combination of ion exchange and
hydrophobic interaction chromatography. No direct affinity chromatography
required. The new process yields a >90% pure protein with no E. coli
RNase carryover.
• Proven Compatibility with RT-PCR Systems: RNasin ® Plus has proven
compatible with the Access and AccessQuick RT-PCR Systems, ImProm-
II Reverse Transcription System and the Reverse Transcription System.
Also proven compatible with TaqMan ® -based RT-PCR Systems.
• Protection During RNA Template Denaturation: Heating mixtures of
RNasin ® Plus and RNase does not lead to reactivation of the RNase at
temperatures even as high as 70°C for 15 minutes. Many RT-PCR
protocols call for RNA template denaturation (e.g., 65–70°C for
5–10 minutes) in the presence of the RT primers prior to full RT reaction
assembly for maximum sensitivity. You can now include RNasin ® Plus at
this step.
• Protection During Higher Temperature RT Reactions: Add RNasin ®
Plus during RT reaction assembly and take the reaction to
temperatures above 50°C with enzymes like the ImProm-II and AMV
Reverse Transcriptases. RNases that may be present will not be reactivated
at the higher temperature.
• Choose Your Configuration: Learn more about our custom options for
this product at: www.promega.com/myway/
Storage Conditions: Store at –20°C.
Protocol Part#
RNasin ® Plus RNase Inhibitor Product Information 9PIN261
For complete and up-to-date product information visit: www.promega.com/catalog
1 2 3 4
Protection from RNase at 70°C. Separate tubes of RNasin ® Plus and RNase
(lanes 1 and 3) were heated to 70°C for 15 minutes. RNasin ® Plus and RNase
were combined and then heated to 70°C for 15 minutes (lanes 2 and 4). To
each set of reactions, either 100ng (lanes 1 and 2) or 10ng (lanes 3 and 4)
of Luciferase Control RNA (Cat.# L4561) were added. The reactions were
held at 37°C for 1 hour, then used in an RT-PCR to amplify the entire 1.8kb
transcript. The gel shows the amplified product from the RT-PCR. All lanes
used 400u of RNasin ® Plus and 1.25μg of a rat liver protein extract (abundant
source of RNase; Sigma Cat.# L-1380) dissolved in water to 0.5μg/μl.
4416TB12_3B
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DNA and RNA Purification
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