2013 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
For Laboratory Use.
10,000 u 40 u/µl N2615 597
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.
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.
Recombinant RNasin ® Ribonuclease
Inhibitor
Product Size Conc. Cat.# Price ($)
Recombinant RNasin ® Ribonuclease 2,500 u 20–40 u/µl N2511 150
Inhibitor
10,000 u 20–40 u/µl N2515 507
For Laboratory Use.
Description: RNases are ubiquitous, cause RNA degradation and can
compromise RNA integrity. Recombinant RNasin ® Inhibitor is a 50kDa protein
that inhibits RNase A family and human placental RNases by noncovalently
binding to RNases in a 1:1 ratio. Recombinant RNasin ® Inhibitor does not
inhibit RNase T1, S1 nuclease, RNase from Aspergillus, RNase H,
RNase ONE Ribonuclease and enzymes for downstream applications
such as GoScript Reverse Transcriptase, AMV/M-MLV reverse transcriptases,
SP6, T7/T3 RNA polymerase, and Taq DNA polymerases. Learn more about our
custom options for this product at: www.promega.com/myway/
Features:
• Inhibits Common Eukaryotic RNases: Carries broad-spectrum RNase
inhibitory properties.
• Compatible: Does not inhibit SP6, T7 or T3 RNA Polymerase; GoScript,
AMV or M-MLV Reverse Transcriptase; or Taq DNA polymerase.
• Broad pH Range (pH 5–8): Offers flexibility in downstream assays.
• Recombinantly Produced: Minimizes chances of human nucleic acid
contamination.
Storage Conditions: Store at –20°C.
Protocol
RNasin ® Ribonuclease Inhibitor, Recombinant Protocol
4416TB12_3B
Part#
9PIN251
6
DNA and RNA Purification
Protocol
RNasin ® Plus RNase Inhibitor Product Information
Part#
9PIN261
Section
Contents
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