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Cat. No.: RNIF-5k (for 5KU)

Cat. No.: RNIF-25k (for 25KU)

Description

RNase If, also known as RNase I‑MBP, is a fusion protein consisting of E. coli RNase I and maltose‑binding protein (MBP), and it retains the same endoribonuclease activity as RNase I. RNase If recognizes and cleaves both single‑stranded RNA (ssRNA) and double‑stranded RNA (dsRNA), generating 5′‑hydroxyl termini and 2′,3′‑cyclic phosphate intermediates, which subsequently hydrolyze to form 3′‑monophosphate nucleotides. The enzymatic activity of RNase If does not require the presence of metal ions, and in the absence of calcium ions, RNase If degrades ssRNA more efficiently than dsRNA. Unlike RNase T1 and RNase A, the activity of RNase If is not inhibited by RNase inhibitor.

A key advantage of RNase If is its heat‑inactivation profile: it can be fully inactivated by heating at 70 °C for 20 minutes, whereas commonly used RNase A and RNase T1 cannot be completely inactivated even when heated to 100 °C.

Applications

Degradation of ssRNA to generate 2′,3′‑cyclic nucleotide intermediates and 3′‑monophosphate nucleotides; removal of RNA from DNA or protein samples; use in ribonuclease protection assays (RPA); RNA analysis and detection.

Source

Recombinant protein expressed in E. coli, encoded by the E. coli RNase I gene fused with an MBP (maltose‑binding protein) tag.

Unit Definition

One unit is defined as the amount of enzyme required to fully digest 1 picomole of synthetic ssRNA 33‑mer in a total reaction volume of 10 µl for 15 minutes in 1× Buffer, as visualized on a 20% acrylamide gel.

Purity

Free of DNase endonuclease and exonuclease activities; contains no ribonucleases other than RNase If.

Enzyme Storage Buffer

10 mM Tris‑HCl (pH 8.0 at 25 °C), 100 mM NaCl, 0.5 mM EDTA, 50% (v/v) glycerol.

Reaction Buffer (10×)

500 mM Tris‑HCl (pH 7.9 at 25 °C), 1 M NaCl, 100 mM MgCl₂, 10 mM DTT.

Inactivation or Inhibition

RNase If can be inactivated by heating at 70 °C for 20 minutes. Complete inactivation can also be achieved by adding 0.1% SDS or by phenol extraction.

Storage

Store at −20 °C; the enzyme remains stable for at least two years. No significant loss of activity is observed after storage at room temperature or 37 °C for three days.

Precautions

• Although the enzyme has been experimentally shown to retain activity after being kept at room temperature or 37 °C for three days, it should still be kept on ice during use and returned to −20 °C immediately after use.
• Because RNA handling is involved, strict RNA‑safe laboratory practices must be followed to avoid RNase contamination.
• All consumables used in the experiment, such as pipette tips and microcentrifuge tubes, must be RNase‑free or treated with DEPC.
• Depending on the specific application, additional reagents such as ultrapure water may be required.
• This product is intended for use by trained professionals for scientific research only. It must not be used for clinical diagnosis or therapy, must not be used in food or drug applications, and must not be stored in residential environments.
• For your safety and health, wear a lab coat and disposable gloves during operation.

Only for research and not intended for treatment of humans or animals

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