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RNase Inhibitor

No Product Name Cat.# Conc Size Price(₩) Add
1 RNase Inhibitor M007S 40 units/µl 2,500 units ₩ 75,000 추가
2 RNase Inhibitor M007M 40 units/µl 5,000 units ₩ 135,000 추가
3 RNase Inhibitor M007L 40 units/µl 12,500 units ₩ 300,000 추가
Product Name Cat.# Size Price(₩) EA Total(₩) 삭제
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Product description
A cDNA encoding human RNase Inhibitor was expressed in E. coli cells, and its recombinant protein was purified to near homogeneity. This product strongly inhibits major nonspecific ribonucleases that include RNase A, B, C, and human placental RNase. However, it does not inhibit sequence-specific ribonucleases or ribonuclease activities associated with DNA or RNA polymerases; these include RNase T1, S1 nuclease, RNase from Aspergillus sp. RNase H, Taq polymerase (Cat.# P025, P050, P225, P250, P725, P750), M-MLV reverse transcriptase (Cat.# M004), SP6, T7 and T3 RNA polymerase.

Characteristics
- Molecular weight : 50 kDa
- Reaction temperature : 25℃-55℃
- Optimal pH range : pH 5.5-9.0
- Heat inactivation : 70℃ for 10 min

Applications
- In vitro transcription
- cDNA synthesis
- RT qPCR
- In vitro translation
- Isolation of mammalian cell fractions that contain mRNAprotein complex
- Separation and identification of specific ribonuclease activities
- Studies of tumor suppression

Quality control
- Purity : >99% on SDS-PAGE
- Endonuclease-free
- Exonuclease-free

Unit definition
One unit of the RNase Inhibitor is the amount of protein required to inhibit 50% of 5 ng RNase A (500 mM Tris- HCl (pH 7.5), 6 mM EDTA, 40 mM DTT, 0.5 mg/ml BSA, 100 ng/ml of RNase A).

Storage buffer
20 mM HEPES-KOH (pH 7.6), 50 mM KCl, 8 mM DTT, 50% (v/v) glycerol

Standard reaction conditions
- Example : in vitro transcription

10X T7 RNA Polymerase buffer

5 μl

T7 RNA Polymerase (50 units/μl)

1 μl

RNase Inhibitor (40 units/μl)

1.25 μl

dNTP mixture (2.5 mM each)

10 μl

DNA template (X μg/ml)

1 μl

Distilled water

up to 50 μl

→ Incubate for 60~120 min at 37


Cautions
- The RNase Inhibitor is active in a wide range of pH.
- At least 1 mM DTT is essential to maintain its activity.
- For efficient protection of RNA in the reaction, add 1 unit of RNase Inhibitor to 1 μl of reaction mixture.


Figure 1. RNA degradation by RNase A can be blocked by RNase inhibitors.
RNaseA (125 ng) was preincubated at 37℃ for 10 min with increasing amounts (8 to 40 units) of RNase inhibitors from Enzynomics (Cat.# M007) or other company (Supplier A). The preincubated mixtures were then added to reaction mixtures containing ~1 μg of in vitro transcribed RNA, followed by incubation at 37℃ for 30 min. The addition of increasing amounts of RNase inhibitors effectively protected RNA from being degraded by RNase A.



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