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| Product Name | Recombinant RNase Inhibitor |
| Catalog No. | IEC-HMM-0053 |
| Description | A recombinant 50 kDa protein that non-covalently binds and inhibits pancreatic-type ribonucleases (RNase A family) including RNase A, RNase B, and RNase C with very high affinity (Ki ~ 3 x 10⁻¹⁰ M). This inhibitor does not affect RNase T1, RNase H, or S1 nuclease activities. |
| Intended Use | Protects RNA from degradation during reverse transcription, in vitro transcription, in vitro translation, and any enzymatic manipulation of RNA where RNase contamination may compromise results. |
| Principle / Technology | Forms a tight 1:1 non-covalent complex with members of the RNase A superfamily through specific protein-protein interactions. Binding is rapid, reversible, and competitive. The inhibitor does not covalently modify the RNase but sterically blocks access to RNA substrates. |
| Detection Method | One unit inhibits 50% of the activity of 5 ng RNase A in a standardized assay using cytidine 2',3'-cyclic monophosphate as substrate under defined buffer conditions. |
| Sample Type | Added as a protective component during RNA handling, reverse transcription, in vitro translation, and RNA labeling reactions. |
| Performance Range / Specifications | 1 U/uL final concentration protects against up to 5 ng/mL RNase A. Typically used at 1 U/uL in most enzymatic reactions. Linear inhibition relationship at recommended concentrations. |
| Sensitivity / LOD | Detectable inhibition of RNase A at inhibitor concentrations as low as 0.01 U/uL. |
| Specificity | Highly specific for RNase A family enzymes. Does not inhibit RNase T1, RNase T2, RNase H, S1 nuclease, DNase I, or other non-pancreatic-type ribonucleases. |
| Reaction Conditions / Protocol | Add to reaction mixture at 1 U/uL final concentration. Compatible with common enzymatic reaction buffers at pH 5.0-9.0. Add before RNA template when setting up reactions. |
| Components / Formulation | Recombinant RNase Inhibitor (40 U/uL) in 20 mM HEPES-KOH (pH 7.6), 50 mM KCl, 8 mM DTT, 50% glycerol. |
| Storage Conditions | Store at -20 °C to -30 °C. Minimize exposure to oxidizing conditions; the inhibitor requires reduced cysteine residues for activity. |
| Shelf Life | 24 months at -20 °C. |
| Package Specifications | 5,000 U (125 uL), 25,000 U (625 uL), 100,000 U (2.5 mL). |
| Product Form | Clear, colorless glycerol-based liquid. Contains DTT which may produce a faint sulfurous odor upon opening. |
| Quality Control | Each lot tested for: inhibitory activity against standard RNase A; absence of contaminating RNase, DNase, and non-specific endonuclease; absence of PCR inhibition; functional validation in reverse transcription reactions measuring RNA integrity. |
| Key Features | Sub-nanomolar binding affinity for RNase A; does not interfere with common molecular biology enzymes; broad pH tolerance; recombinant source eliminates animal-derived material concerns. |
| Purity | >= 95% by SDS-PAGE. |
| Concentration | 40 U/uL standard. Custom concentrations (20, 60, 80 U/uL) available. |
| Activity / Unit Definition | 1 unit inhibits 50% activity of 5 ng RNase A under standardized assay conditions. |
| Molecular Weight | ~50 kDa (monomer). Active as monomer; each molecule binds one RNase molecule. |
| Source / Origin | Recombinant protein produced in E. coli. Gene derived from human placental RNase inhibitor sequence, codon-optimized for bacterial expression. |
| pH Range / Optimal pH | Active from pH 5.0 to 9.0 with maximum inhibition at pH 7.0-8.0. DTT (1-10 mM) required to maintain reducing environment for cysteine sulfhydryl groups. |
| Shipping Conditions | Shipped with cold packs to maintain below 10 °C. Avoid freezing at temperatures above -20 °C. |
| Expiration Date / Stability | 24 months at -20 °C. Activity declines in the absence of reducing agents. Ensure DTT concentration is maintained if diluting the stock solution. |
| Regulatory / Compliance | ISO 9001 certified production. For research use. Recombinant source suitable for IVD applications with appropriate regulatory documentation. |
| Compatibility | Fully compatible with M-MLV and AMV reverse transcriptase reactions, T7/SP6/T3 in vitro transcription, in vitro translation (rabbit reticulocyte, wheat germ), and RNA labeling protocols. Do not use in reactions containing >8 M urea or strong denaturants. |
| Recommended Buffer System | Compatible with common Tris, HEPES, and phosphate buffers. Requires 1-10 mM DTT or beta-mercaptoethanol for stability. Avoid buffers containing p-chloromercuribenzoate or other sulfhydryl-modifying agents. |
| Application Notes / Precautions | Add inhibitor to reaction master mix before adding RNA. For long incubations (>2 h), supplement with additional DTT to 5 mM. The inhibitor is inactivated by heating above 65 °C, so add it after any heat-denaturation step when working with RNA. |
| Batch-to-Batch Consistency | Inhibition titer consistent within +/- 10% between lots. RNase-free certification verified for every production batch. |
For research use only, not for clinical use.
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