Murine RNase inhibitor GMP-grade

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Cat Number

PIPB-0798

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Product Detail Description Scientific Support Customer Q&A

Storage

Store at -20 ℃

Molecular Weight

50 kDa

Standard

Facility GMP

Qualification

DMF

General Description

Murine RNase Inhibitor is a recombinant, GMP grade, highly purified, stabilized protein expressed and purified in E. coli and supplied in liquid form. Oxidation sensitive human or porcine RNase inhibitors contain cysteine residues that are easily oxidized. Murine RNase inhibitor has none of the oxidation sensitive cysteine residues, making this inhibitor much more oxidation resistant.

Mechanism of Action

Murine RNase Inhibitor is a strong, non-competitive inhibitor that binds non-covalently to form a stable 1: 1 stoichiometric complex with the target enzyme in a highly specific manner. This high affinity inhibitor inactivates the specific ribonucleases, RNases A, B and C, thus protecting valuable RNA from degradation. Murine RNase Inhibitor does not inhibit common polymerases (e.g. Taq DNA Polymerase or Reverse Transcriptases).

Application

Murine RNase Inhibitor is typically used in sensitive molecular biology applications to protect RNA integrity. It is necessary to protect RNA during RT-PCR and RT-qPCR, cDNA synthesis, in vitro transcription and translation assays and wherever highly sensitive reactions are performed or the structural and functional integrity of RNA is required. It has excellent antioxidant activity and is stable at low DTT concentrations.

seCLIP is a simplified, single-end variant of eCLIP for transcript-wide mapping of RNA-binding-protein (RBP) targets with nucleotide resolution. Redesigning the adapters, it obviates the expensive paired-end sequencing and retains quantitative precision and motif recovery.
Key protocol notes are noted: (1) supplementing with murine RNase inhibitor during the initial lysis step for RNase-rich tissues to avoid over-fragmentation; (2) switching to low-background lots of nitrocellulose membranes in case of contamination; and (3) using qPCR-guided PCR cycles to limit duplication. The workflow (UV crosslinking, RNase I trimming, immunoprecipitation, on-bead RNA/DNA ligation, PAGE isolation and a high-efficiency library construction) provides >1000-fold higher yield compared to older versions of CLIP, allowing for robust profiling of RBPs from low-input material at lower cost.

Fig. 1 Murine RNase inhibitor is used to avoid RNA over-fragmentation. (Van Nostrand E L.; et al. 2017)

References

Van Nostrand E L, et al. Robust, cost-effective profiling of RNA binding protein targets with single-end enhanced crosslinking and immunoprecipitation (seCLIP). mRNA Processing: Methods and Protocols. New York, NY: Springer New York, 2017: 177-200.

How does Murine RNase inhibitor work?

It binds RNase to form an inactive complex, specifically inhibiting RNase A, B, and C.

What is the source of your Murine RNase inhibitor?

Recombinant, mouse-derived protein expressed and purified from E. coli.

Is Murine RNase inhibitor tested for contaminants?

Yes, it is tested to be free of DNase and RNase activity.

What applications is Murine RNase inhibitor recommended for?

RT-PCR, cDNA synthesis, in vitro transcription, enzymatic RNA labeling and RNA integrity applications.