RNase Inhibitor, IC-5226
Product Introduction
This product is a recombinant murine RNase inhibitor expressed in an E. coli expression system. RNase inhibitor is a 50 kDa protein that binds non-covalently to RNase enzymes, rendering them inactive and thereby protecting RNA from degradation during molecular biology procedures. The inhibitor specifically binds to RNase A, B, and C through a non-competitive inhibition mechanism, effectively suppressing their enzymatic activity. The inhibition is reversible: upon addition of urea or sulfhydryl reagents, the inhibitor is inactivated and RNase A, B, and C regain activity. This product does not inhibit RNase 1, RNase T1, S1 nuclease, RNase H, or RNase derived from Aspergillus species. Importantly, it exhibits no inhibitory effect on Taq DNA polymerase, AMV or M-MLV reverse transcriptases, or phage RNA polymerases (SP6, T7, or T3), making it fully compatible with a wide range of downstream enzymatic reactions. This formulation is cysteine-free, conferring enhanced oxidation resistance and improved stability at low DTT concentrations (1 mM). The RNase inhibitor is an essential reagent for any RNA-based application requiring preservation of RNA integrity.
Product Features
1. Broad RNase Inhibition: Non-competitively and specifically inhibits RNase A, B, and C, providing robust protection of RNA in enzymatic reactions.
2. Compatible with Downstream Enzymes: Does not inhibit Taq DNA polymerase, AMV/M-MLV reverse transcriptases, or SP6/T7/T3 phage RNA polymerases, ensuring seamless integration into cDNA synthesis, RT-PCR, and in vitro transcription workflows.
3. Cysteine-Free Formulation: Enhanced oxidation resistance and improved stability at low DTT concentrations (1 mM), reducing potential interference with disulfide bond-dependent reactions.
4. Reversible Inhibition: Inhibition can be reversed by urea or sulfhydryl reagents, enabling controlled RNase reactivation if required for specific applications.
5. High Specificity: Does not inhibit RNase 1, RNase T1, S1 nuclease, RNase H, or Aspergillus-derived RNases, providing predictable and reliable specificity.
Specifications
Size: Available in 2,000 U (2KU)
Source: Recombinant murine RNase inhibitor expressed in E. coli
Molecular Weight: 50 kDa
Definition: One unit is defined as the amount of enzyme required to inhibit the activity of 5 ng of RNase A by 50%.
Inhibition Mechanism: Non-competitive, non-covalent binding to RNase A, B, and C
Key Exclusions: Does not inhibit RNase 1, RNase T1, S1 nuclease, RNase H, Aspergillus-derived RNases
Enzyme Compatibility: Compatible with Taq DNA polymerase, AMV/M-MLV reverse transcriptases, SP6/T7/T3 phage RNA polymerases
Applications:
1. cDNA synthesis
2. RT-PCR
3. In vitro transcription/translation
4. RNase-protected RNA labeling reactions
5. Other applications requiring RNA integrity
Storage and Stability
Storage Conditions: Store at -20¡ãC. Avoid repeated freeze-thaw cycles.
Shelf Life: The product is stable for 12 months from the date of manufacture when stored as directed.
Protocol (For Reference Only)
Important: Work in an RNase-free environment. Use RNase-free pipette tips, tubes, and reagents. Wear gloves and change them frequently to prevent RNase contamination from skin. Add RNase inhibitor as the final component to reaction mixtures.
1. cDNA Synthesis: Add 1 µL of RNase Inhibitor (20-40 U) per 20 µL reverse transcription reaction. Mix gently by pipetting; do not vortex. The inhibitor is compatible with both AMV and M-MLV reverse transcriptases.
2. RT-PCR: For two-step RT-PCR, add 1 µL of RNase Inhibitor per 20 µL reverse transcription reaction. For one-step RT-PCR, add 0.5-1 µL per 25-50 µL reaction. The inhibitor does not interfere with Taq DNA polymerase activity.
3. In Vitro Transcription: Add 1-2 µL of RNase Inhibitor (20-40 U) per 20 µL transcription reaction containing SP6, T7, or T3 RNA polymerase. The inhibitor protects the RNA transcript from degradation during the reaction.
4. In Vitro Translation: Add 1 µL of RNase Inhibitor (20-40 U) per 25-50 µL translation reaction mixture to protect mRNA templates during protein synthesis.
5. RNA Labeling and Modification: For RNA end-labeling, ligation, or other enzymatic modification reactions, add 1 µL of RNase Inhibitor per 20-50 µL reaction to prevent RNA degradation during incubation.
Precautions
1. Use RNase-free techniques throughout. The RNase inhibitor is not a substitute for proper RNase-free laboratory practice.
2. Add the RNase inhibitor as the final component to reaction mixtures to ensure immediate inhibition of any contaminating RNases.
3. Avoid repeated freeze-thaw cycles; aliquot into single-use volumes upon receipt.
4. This product does not inhibit all RNases; it is specific for RNase A, B, and C. Consider the source of potential RNase contamination when designing experiments.
5. For research use only. Not for use in diagnostic or therapeutic procedures.
FAQ (Simplified)
Q1: What RNases does this inhibitor target?
A1: This RNase inhibitor specifically and non-competitively inhibits RNase A, B, and C. It does not inhibit RNase 1, RNase T1, S1 nuclease, RNase H, or RNases derived from Aspergillus species.
Q2: Is this inhibitor compatible with reverse transcriptases and DNA polymerases?
A2: Yes. This RNase inhibitor does not inhibit AMV reverse transcriptase, M-MLV reverse transcriptase, Taq DNA polymerase, or phage RNA polymerases (SP6, T7, T3). It is fully compatible with all standard molecular biology enzymatic reactions.
Q3: How is this product different from other RNase inhibitors?
A3: This recombinant murine RNase inhibitor is cysteine-free, which enhances its oxidation resistance and improves stability at low DTT concentrations (1 mM). This formulation characteristic reduces potential interference with disulfide bond-dependent enzymatic reactions.
Q4: How should I store this product?
A4: Store at -20¡ãC. Avoid repeated freeze-thaw cycles by aliquoting into single-use volumes upon receipt. The inhibitor maintains full activity under recommended storage conditions for 12 months from the date of manufacture.
Disclaimer
1. For Research Use Only. Not for use in diagnostic or therapeutic procedures.
2. Due to the variable nature of biological research, optimization of inhibitor concentration is recommended for specific experimental systems.
3. This warranty is limited to the replacement of the product. The manufacturer assumes no liability for incidental or consequential damages, including loss of samples or data.
4. Wear appropriate protective clothing and gloves when handling this product. Use RNase-free consumables and maintain RNase-free technique.
Ordering Information
Catalog Number: IC-5226
Product Name: RNase Inhibitor
Size: 2,000 U (2 KU)
Price:
2,000 U: CNY ¥1080.00 / USD $108.00 / EUR €132.00 / JPY ¥19800.00
2023 Version
