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2012 Promega catalogue

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Life<br />

Science<br />

Catalog<br />

<strong>2012</strong><br />

Worldwide Contact List<br />

Section<br />

Contents<br />

Table of<br />

Contents<br />

Cell Signaling<br />

Ribonuclease Inhibitors<br />

96<br />

RNasin ® Plus RNase Inhibitor<br />

Product Size Conc. Cat.# Price ($)<br />

RNasin ® Plus RNase Inhibitor 2,500 u 40 u/µl N2611 173.00<br />

For Laboratory Use.<br />

10,000 u 40 u/µl N2615 597.00<br />

Description: RNasin ® Plus RNase Inhibitor is a recombinant mammalian<br />

RNase inhibitor that is expressed as a soluble protein in E. coli, allowing easy<br />

purification through a combination of ion exchange and hydrophobic interaction<br />

chromatography. The protein is capable of inhibiting eukaryotic RNases (e.g.,<br />

RNase A and RNase B) similarly to human placental RNase inhibitor. RNasin ®<br />

Plus RNase Inhibitor is tested in RT-PCR and is compatible with enzymes such<br />

as AMV, M-MLV and ImProm-II Reverse Transcriptases or Taq and Tfl DNA<br />

Polymerases. RNasin ® Plus RNase Inhibitor also is tested and compatible with<br />

quantitative, real-time RT-PCR in a TaqMan ® assay.<br />

The inhibitor offers increased resistance to oxidation over the human version of<br />

the protein. Two cysteines in the human protein have been identified as<br />

especially sensitive to oxidation and react by forming a disulfide bond that can<br />

block the active site of the inhibitor. RNasin ® Plus, through natural amino acid<br />

diversity, lacks the ability to form this site-blocking disulfide. In addition, the<br />

new protein has characteristics never before realized, including continued<br />

inhibition of RNases above 50°C. Heating solutions of RNasin ® Plus and RNase<br />

followed by cooling does not result in the reappearance of RNase activity—<br />

even when the solution is heated above the denaturation temperature of the<br />

RNasin ® Plus protein alone. This allows RNasin ® Plus to protect RNA species<br />

prior to, during and after heating, even at temperatures normally used during<br />

first-strand DNA synthesis in RT-PCR. We have taken solutions up to 70°C for<br />

15 minutes and did not see RNase reactivation.<br />

Features:<br />

• Improved Resistance to Oxidation: Due to natural amino acid diversity,<br />

RNasin ® Plus lacks the capability to form the active site-blocking disulfide<br />

bond that can form in the human protein under oxidative conditions.<br />

• Improved Purification: RNasin ® Plus is expressed by E. coli as a soluble<br />

protein, allowing easy purification by a combination of ion exchange and<br />

hydrophobic interaction chromatography. No direct affinity chromatography<br />

required. The new process yields a >90% pure protein with no E. coli<br />

RNase carryover.<br />

• Proven Compatibility with RT-PCR Systems: RNasin ® Plus has proven<br />

compatible with the Access and AccessQuick RT-PCR Systems, ImProm-<br />

II Reverse Transcription System and the Reverse Transcription System.<br />

Also proven compatible with TaqMan ® -based RT-PCR Systems.<br />

• Protection During RNA Template Denaturation: Heating mixtures of<br />

RNasin ® Plus and RNase does not lead to reactivation of the RNase at<br />

temperatures even as high as 70°C for 15 minutes. Many RT-PCR<br />

protocols call for RNA template denaturation (e.g., 65–70°C for<br />

5–10 minutes) in the presence of the RT primers prior to full RT reaction<br />

assembly for maximum sensitivity. You can now include RNasin ® Plus at<br />

this step.<br />

• Protection During Higher Temperature RT Reactions: Add RNasin ®<br />

Plus during RT reaction assembly and take the reaction to<br />

temperatures above 50°C with enzymes like the ImProm-II and AMV<br />

Reverse Transcriptases. RNases that may be present will not be reactivated<br />

at the higher temperature.<br />

• Choose Your Configuration: Learn more about our custom options for<br />

this product at: www.promega.com/myway/<br />

Storage Conditions: Store at –20°C.<br />

Protocol Part#<br />

RNasin ® Plus RNase Inhibitor Product Information<br />

9PIN261<br />

RNasin ® Ribonuclease Inhibitors<br />

Product Size Conc. Cat.# Price ($)<br />

RNasin ® Ribonuclease Inhibitor 2,500 u 20–40 u/µl N2111 Pls. Enq.<br />

10,000 u 20–40 u/µl N2115 Pls. Enq.<br />

Recombinant RNasin ®<br />

2,500 u 20–40 u/µl N2511 150.00<br />

Ribonuclease Inhibitor<br />

10,000 u 20–40 u/µl N2515 507.00<br />

N2511, N2515 For Laboratory Use. N2111, N2115 For Research Use Only. Not for Use in<br />

Diagnostic Procedures.<br />

Description: Natural and Recombinant RNasin ® Ribonuclease Inhibitors<br />

have broad-spectrum RNase inhibitory properties, including the inhibition<br />

of eukaryotic RNases of the neutral type. The 50kDa protein exerts its<br />

inhibitory effect by noncovalently binding to RNases in a 1:1 ratio. The K i value<br />

for the binding of RNasin ® Ribonuclease Inhibitor to RNase (e.g., RNase A) is<br />

approximately 10 –14 M. <strong>Promega</strong> offers two different preparations: Recombinant<br />

RNasin ® Ribonuclease Inhibitor and Natural RNasin ® Ribonuclease Inhibitor.<br />

Both products are purified using a combination of ion exchange and affinity<br />

chromatography.<br />

Features:<br />

• Inhibits Common Eukaryotic RNases: RNasin ® Ribonuclease Inhibitor<br />

has broad-spectrum RNase inhibitory properties, including RNase A, RNase<br />

B, RNase C and human placental RNase. Does not inhibit RNase T1, S1<br />

nuclease, RNase from Aspergillus, RNase H, RNase ONE Ribonuclease.<br />

• Compatibility: RNasin ® Ribonuclease Inhibitor does not inhibit SP6, T7 or<br />

T3 RNA Polymerase; ImProm-II, AMV or M-MLV Reverse Transcriptase;<br />

or Taq DNA polymerase.<br />

• Broad pH Range: Active over a broad pH range (pH 5–8).<br />

• Choose Your Configuration: Learn more about our custom options for<br />

this product at: www.promega.com/myway/<br />

Storage Conditions: Store at –20°C.<br />

Protocol Part#<br />

RNasin ® Ribonuclease Inhibitor Product Information<br />

9PIN211<br />

RNasin ® Ribonuclease Inhibitor, Recombinant Protocol<br />

9PIN251<br />

See figure on next page for more information.<br />

For complete and up-to-date product information visit: www.promega.com/catalog

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