RIBOPROTECT Hu RNase Inhibitor (RT35)

The RIBOPROTECT Hu RNase Inhibitor is a 50 kDa recombinant human placental protein expressed in Escherichia coli. It inhibits ribonuclease (RNase) activity of common eukaryotic enzymes such as RNase A, RNase B, RNase C, and protects RNA integrity from degradation. RIBOPROTECT Hu 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. It is compatible with DNA Polymerases and AMV or M-MuLV Reverse Transcriptases.
Available Options:
SKU Unit
RIBOPROTECT Hu RNase Inhibitor RT35 RT35-020 2000 U ( 40U/μl ) Ask for an offer
RIBOPROTECT Hu RNase Inhibitor RT35 RT35-100 10 000 U ( 40U/μl ) Ask for an offer

RIBOPROTECT RNase Inhibitor – Maximum RNA protection

RIBOPROTECT Hu RNase Inhibitor is the right choice to overcome the challenges coming with the presence of ubiquitous RNases, commonly found in skin, dust, reagents, and biological samples.

The RIBOPROTECT Hu RNase Inhibitor is a 50 kDa recombinant human placental protein expressed in Escherichia coli. It inhibits ribonuclease (RNase) activity of common eukaryotic enzymes such as RNase A, RNase B, RNase C. by non-covalent binding in a 1:1 ratio. RIBOPROTECT 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. Such inhibition effectively helps to maintain the RNA integrity and allows to obtain the appropriate quantity and quality of RNA.

RIBOPROTECT Hu is intended for use in applications where the presence of RNases may cause 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. RIBOPROTECT Hu shows no activity towards RNase 1, RNase T1, RNase T2, S1 nuclease, and RNase H.

It is compatible with DNA Polymerases and AMV or M-MuLV Reverse Transcriptases.

Features and advantages

  • Completely inhibits RNase A, B, and C activity
  • No influence on the polymerase or reverse transcriptase activity
  • Free of DNase and RNase activity
  • Retains full stability at 37°C for at least 4 weeks
  • Stable up to 58°C and at min. 0.5 – 1 mM DTT concentration ranges
  • Active in diverse reaction conditions and in various buffers
  • Active over a broad pH range (pH 5.5 – 9.0)
  • Compatible with the TRANSCRIPTME Reverse Transcriptase (cat. no. RT32)

Applications

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

Unit definition

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

Stability during shipment

RIBOPROTECT displays unaffected activity for 4 weeks when incubated at 37 ̊C. Such a high unchanged activity in challenged temperature conditions is a guarantee of stability during uncontrolled shipment incidents.

RNase Inhibitor stability

RNase Inhibitor – Batch-to-batch consistency

Our customers’ positive feedback is our best proof of reliability. The closeness in activity in the following three RIBOPROTECT batches (purple columns), coming from three different product purifications, shows very high batch-to-batch consistency. The grey column represents activity from a competitive RNase Inhibitor.

RNase Inhibitor Batch-to-batch consistency

The activity (U/μl) of RIBOPROTECT from three separate batches (purple columns) and a competitive RNase Inhibitor (grey column) were tested by BLIRT’s customer.

 

The efficiency comparison of RIBOPROTECT and competitive RNase Inhibitors

We compared the inhibition efficiency of the RNA degradation by RIBOPROTECT and two broadly used RNase Inhibitors from the top market leaders (marked here as RNase Inhibitor T and RNase Inhibitor P). We incubated 1 μg RNA (kidney cancer cell lines, 786-O) with 5 ng RNase A (BLIRT) at 37°C for 15 min with various RNase Inhibitor concentrations: 20, and 40U in a 20 μl reaction.

RIBOPROTECT displays very high RNA protection (RIN: 9.667±0.240). For the recommended concentration of 40U, no differences were found between RIBOPROTECT and RNase InhibitorP, while RNase Inhibitor T showed lower protection (RIN: 9.678 vs. 8.733, p <0.05). Our results display that both RNase Inhibitor T and P are less efficient than RIBOPROTECT at 20U per 20 μl reaction (p <0.0001).

 

RNAse Inhibitor efficiency

RIBOPROTECT is the most efficient among all three tested RNase Inhibitors. Its equally high efficiency at 20-40U/20 μl reaction, which BLIRT recommends for most applications proves its high competitiveness among other RNase Inhibitors.

RNase Inhibitor efficiency

RIBOPROTECT Hu RNase Inhibitor (RT35) - Product Specification

RIBOPROTECT Hu RNase Inhibitor (RT35) - Manual

RIBOPROTECT Hu RNase Inhibitor (RT35) - Flyer

RIBOPROTECT Hu RNase Inhibitor (RT35) - Complete portfolio for RNA workflow

RIBOPROTECT Hu RNase Inhibitor – Troubleshooting

Usage

  • The optimal final concentration of the RIBOPROTECT 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 RIBOPROTECT Hu for a final sample volume of 20 μl.
  • For the optimal RIBOPROTECT Hu activity, a final DTT concentration of 0.5 – 1 mM is essential.
  • During assembly of a reaction, RIBOPROTECT Hu should be added before other components that are possible sources of RNase contamination.
  • Using RIBOPROTECT Hu does not exclude RNase H treatment after amplification of the first strand cDNA.

Storage buffer

20 mM HEPES-KOH (pH 7.6); 50 mM KCl; 8 mM reducing agent; 50% (v/v) glycerol

Storage conditions

Store at -20°C in a freezer without a defrost cycle.

Quality control

The absence of Endonuclease, Exonuclease, RNase and latent RNase activities has been confirmed using the relevant procedures. The purity is >90% as judged by SDS-polyacrylamide gels.

RIBOPROTECT Hu RNase Inhibitor (RT35) - MSDS


RIBOPROTECT Hu RNase Inhibitor (RT35) - CoA

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