RNase Inhibitor Hu

For maximum RNA protection where intact RNA is required

S_1353_3_LS_OEM_RNase_Inhibitor_Hu_2000_U
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RNase Inhibitor Hu (2000 U)

Cat. No. / ID:  RT35-020

Storage temperature: -20°C without a defrost cycle
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$76.00
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Quantity
2000 U
2500 U
10000 U
20000 U
40000 U
400000 U
1600000 U
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The RNase Inhibitor Hu is intended for molecular biology applications. This product is neither intended for the diagnosis, prevention or treatment of a disease, nor has it been validated for such use either alone or in combination with other products.
Need bulk, customized or optimized products for commercial purposes? We also offer support with logistics, compliance and more. Reach out to cooperate with QIAGEN Strategic Partnerships & OEM

Features

  • Completely inhibits RNase A, RNase B, and RNase C activity
  • Does not influence any polymerase or reverse transcriptase activity
  • Retains full stability at 37°C for at least 4 weeks with no loss of activity
  • Active in diverse reaction conditions and in various buffers
  • Free of DNase and RNase activity

Product Details

RNase Inhibitor Hu is a thermally resistant, 50 kDa recombinant human placental protein expressed in Escherichia coli that inhibits RNase activity of common eukaryotic enzymes such as RNase A, RNase B, RNase C.


It is supplied with 20 mM Hepes-KOH (pH 7.6), 50 mM KCl, 8 mM reducing agent, 50% (v/v) glycerol. It is stable up to 58°C and at a minimum 0.5–1 mM DTT concentration ranges.


One unit is defined as the amount of enzyme required to inhibit the activity of 5 ng RNase A by 50%.

 

Performance

Assay Specification
Purity >90%
RNase contamination None detected
Latent RNase contamination None detected
Exonuclease contamination None detected
Endonuclease contamination None detected

 

 

Principle

RNase Inhibitor Hu inhibits RNases by non-covalent binding in a 1:1 ratio with high-affinity protein-protein interaction, forming one of the tightest known biomolecular complexes. This activity effectively helps in the maintenance of RNA integrity and in obtaining the appropriate quantity and quality of RNA.


The enzyme is used in applications where the presence of RNases causes a substantial hazard in receiving good quality, trustworthy, and reproducible data, e.g., in RNA isolation, cDNA synthesis, RT-PCR, in vitro transcription and translation, or RNase-free monoclonal antibody preparation [1-6]. It is compatible with numerous reagents, including DNA polymerases and AMV or M-MuLV Reverse Transcriptase, and its addition into the reaction does not influence the efficiency of other components.

 

Procedure

Quality Control


Protein purity is determined using assay by SDS-PAGE electrophoresis resulting in >90% purity.


RNase contamination is evaluated by assessing RNase activity through incubation of 1 µg of RNA with a minimum of 200 U of the RNase Inhibitor for 1 hour at 37°C. Results are visualized on an ethidium bromide-stained agarose gel.


Latent RNase contamination is evaluated by assessing latent RNase activity through incubation of 1 µg of RNA with a minimum of 200 U of heat-inactivated RNase Inhibitor for 1 hour at 37°C. Results are visualized on an ethidium bromide-stained agarose gel.


Endonuclease activity is determined by incubating 1 µg of supercoiled plasmid DNA with a minimum of 200 U of the RNase Inhibitor for 2 hours at 37°C. Results are visualized on an ethidium bromide-stained agarose gel.
Exonuclease activity is determined by incubating 1 µg of digested plasmid DNA with a minimum of 200 U of the RNase Inhibitor Lyo-ready for 2 hours at 37°C. Results are visualized on an ethidium bromide-stained agarose gel.

Usage

  • The optimal final concentration of the RNase Inhibitor Hu in a reaction depends on the level of RNase contamination, the incubation time and the compounds present in the reaction mixture. It falls within a range of 1–2 U/μL.
  • For a standard reverse transcription reaction, use 1 μL (40 U) of the RNase Inhibitor Hu for a final sample volume of 20 μL.
  • A final DTT concentration of 0.5–1 mM is essential for optimal activity.
  • During the reaction assembly, the enzyme should be added before other components that are possible sources of RNase contamination.
  • Using RNase Inhibitor Hu does not exclude RNase H treatment after amplification of the first strand cDNA.

 

Applications

This is used for applications to overcome the challenges in inhibiting the presence of ubiquitous RNases, commonly found in skin, dust, reagents, and biological samples, such as:

  • RNA-related molecular diagnostics
  • RNA isolation and purification
  • cDNA synthesis, RT-PCR, RT-qPCR
  • in vitro transcription and translation
  • RNase-free monoclonal antibody preparation

 

Resources

Quick-Start Protocols (1)
Safety Data Sheets (1)
Certificates of Analysis (1)