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| Product Name | Recombinant RNase Inhibitor (Murine-Origin, Oxidation-Resistant) |
| Catalog No. | IEC-HMM-0070 |
| Description | A recombinant murine-origin ribonuclease inhibitor protein produced in E. coli, engineered with cysteine-to-serine substitutions to eliminate the oxidation sensitivity of wild-type placental RNase inhibitor. This variant retains full inhibitory activity against RNase A, RNase B, and RNase C but does not require DTT in the storage and reaction buffer. The DTT-independent formulation makes it compatible with applications where reducing agents interfere, such as maleimide-based protein labeling. |
| Intended Use | Protection of RNA from ribonuclease degradation during reverse transcription, in vitro transcription, and RNA handling procedures where DTT must be avoided. |
| Principle / Technology | Tight non-competitive binding inhibition of pancreatic-type ribonucleases |
| Detection Method | RNA integrity assessment by gel electrophoresis or Bioanalyzer after incubation with RNase in presence of inhibitor |
| Sample Type | RNA in aqueous solution; compatible with RT-PCR, IVT, and RNA storage workflows |
| Performance Range / Specifications | Inhibitory concentration: 1 unit inhibits 5 ng RNase A by 50% in a standard RNase activity assay; 1 unit per 20 µL reaction typically sufficient for RT applications |
| Sensitivity / LOD | Complete protection of 1 µg RNA from 10 pg RNase A for 1 hr at 37 °C at recommended concentration |
| Specificity | Inhibits RNase A, B, C; does not inhibit RNase H, RNase T1, S1 nuclease, or RNase III family enzymes |
| Reaction Conditions / Protocol | Add 1 µL (40 U) per 20 µL RT or IVT reaction; no DTT or other reducing agent required |
| Components / Formulation | Recombinant RNase Inhibitor (40 U/µL in storage buffer without DTT), 5× RT-compatible buffer |
| Storage Conditions | –20 °C; stable through 10 freeze-thaw cycles |
| Shelf Life | 24 months at –20 °C |
| Package Specifications | 2000 U, 10000 U |
| Product Form | Liquid enzyme in non-reducing storage buffer |
| Quality Control | RNase inhibition verified by protection of in vitro transcribed RNA from RNase A challenge; absence of DNase and non-specific RNase activity confirmed |
| Key Features | DTT-independent formulation compatible with oxidized protein labeling and thiol-reactive conjugation chemistries |
| Purity | ≥95% by SDS-PAGE for enzymes; ≥98% by HPLC for coenzymes |
| Concentration | Enzymes at specified U/µL; coenzymes at specified mM concentration |
| Activity / Unit Definition | One unit incorporates 10 nmol dNTP into acid-insoluble form in 30 min at specified temperature (polymerases); other enzyme units per product specification |
| Molecular Weight | As specified per enzyme: Taq ~94 kDa; M-MLV RT ~75 kDa; T4 ligase ~55 kDa; coenzymes as labeled |
| Source / Origin | Recombinant, expressed in E. coli; purified by multi-step chromatography |
| pH Range / Optimal pH | pH 7.5–8.5 for polymerase activity; pH 7.5–8.0 for ligation; pH 7.5–8.0 for transcription |
| Shipping Conditions | Dry ice for enzyme shipments; cold packs for coenzyme solutions |
| Expiration Date / Stability | 24 months at –20 °C for enzymes; 12–36 months for coenzymes per product type |
| Regulatory / Compliance | Research use; ISO 9001 manufacturing; GMP-grade options available for diagnostic manufacturing |
| Compatibility | Compatible with standard PCR, RT-PCR, isothermal amplification, and molecular cloning workflows |
| Recommended Buffer System | Tris-HCl, KCl, MgCl₂-based buffers; glycerol-containing storage buffers |
| Application Notes / Precautions | Aliquot enzymes to avoid repeated freeze-thaw; use dedicated nuclease-free workspace; verify enzyme activity for critical applications |
| Batch-to-Batch Consistency | Enzyme specific activity within ±15% of reference lot; functional testing per lot |
For research use only, not for clinical use.
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