Highly Pure Ribonuclease Inhibitor Solution - RNase Inhibitor 40U/μl, Optimal for RNA Protection in cDNA Synthesis and Molecular Biology Applications

Product Description

Introducing our Highly Pure Ribonuclease Inhibitor Solution, a premium quality reagent featuring RNase Inhibitor at an ultra-concentrated 40 Units/μl. This solution is specifically engineered to provide maximum protection against ribonucleases during RNA handling in various molecular biology procedures.

Key Features

• Exceptional Purity: The Ribonuclease Inhibitor is recombinantly produced and stringently purified to guarantee a high degree of purity, ensuring no interference with your experiments.

• Ultra-Concentrated Formula: Supplied at 40U/μl concentration, this inhibitor allows for efficient use in demanding applications where even trace amounts of RNase can compromise results.

• Broad-Spectrum Activity: The inhibitor effectively neutralizes a wide array of RNases A, B, C, and other endo- and exoribonucleases, safeguarding your RNA samples from degradation.

• Stable and Active: Formulated for stability across a range of temperatures and pH conditions, the inhibitor remains active during storage and throughout various experimental protocols.

• Compatibility: Ideal for use with automated systems and hand-prepared reactions, it is fully compatible with buffers and enzymes commonly used in cDNA synthesis and nucleic acid manipulation techniques.

Applications

• cDNA Synthesis: Essential for preventing RNase contamination during reverse transcription reactions, thus improving the yield and integrity of synthesized cDNA.

• RNA Isolation and Purification: Vital component in RNA extraction kits and purification workflows to maintain RNA integrity before and after isolation.

• Northern Blotting: Ensures high-quality RNA transfer and hybridization by inhibiting RNase activity that could degrade RNA on membranes.

• qPCR and RT-qPCR: Protects RNA templates in quantitative PCR and reverse transcription quantitative PCR assays, enhancing sensitivity and specificity.

• RNA-Protein Interactions Studies: Preserves RNA molecules during co-immunoprecipitation (RIP) and other RNA-protein interaction experiments.

Source

E. coli strain expressing a recombinant clone that carries Ribonuclease Inhibitor gene from human placenta.

Concentration

40 U/μl.

Unit Definition

One unit is defined as the amount of Recombinant Ribonuclease Inhibitor required inhibit the activity of 5ng of ribonuclease A by 50%.

Activity is measured by the inhibition of hydrolysis of cytidine 2’3’-cyclic monophosphate by ribonuclease A．

Storage Buffer

20mM HEPES-KOH(PH7.6), 50mM KCl, 8mM DTT, 50%(v/v) glycerol

Storage

Store at -20°C

Quality Control

Absence of Endonuclease

1μg of Lambda DNA is incubated with 200 units of Ribonuclease Inhibitor for 16 hours at 37°C.

Following incubation, Lambda DNA is visualized as intact on an ethidium bromide-stained agarose gel to verify the absence of visible Endonuclease.

Absence of Nickase

1μg of Type I supercoiled pBR322 is incubated with 200 units of Ribonuclease Inhibitor for 4 hours at 37°C.

Following incubation, the supercoiled DNA is visualized on an ethidium bromide-stained agarose gel to verify the absence of visible nicking or cutting.

Absence of Exonuclease

1μg of Lambda DNA/HindIII Markers is incubated with 200 units of Ribonuclease Inhibitor for 16 hours at 37°C.

Following incubation, Lambda DNA/HindIII Markers is separated by 1% agarose gel and stained with ethidium bromide. Markers remain as intact bands without smearing.

Absence of RNase and latent RNase

To test for the presence of RNase activity, 1ug of RNA is incubated with 200 units of Ribonuclease Inhibitor for 4 hours at 37°C.

Following incubation, the RNA is visualized as intact band on an ethidium bromide-stained agarose gel to verify the absence of visible RNase.

To test for the presence of latent RNase activity, 200units of Ribonuclease Inhibitor is heat-denatured at 70°C for 15minutes and incubated with 1ug of RNA for 4 hours at 37°C.

Following incubation, the RNA is visualized as intact band on an ethidium bromide-stained agarose gel to verify the absence of visible RNase.

Physical Purity

The purity is ≥95% as judged by SDS-polyacrylamide gel with Coomassie blue staining.

Usage Note

Ribonuclease Inhibitor is active over a broad pH range (pH5.5-9). The standard RT and in vitro transcription uses Ribonuclease Inhibitor at a final concentration of 1u/ul.