Tanım
RNase inhibitor is a specific ribonuclease (RNase) inhibitor found in the human placenta. Its principle of specifically inactivating RNase is: binding to RNase via non-covalent bonds to form a complex, thereby inactivating the RNase.
This product is a recombinant murine RNase inhibitor expressed and purified in a soluble form in E. coli, capable of broadly inhibiting various types of RNase (RNase A, B, C). Verified by RT-PCR and RT-qPCR assays, this product is compatible with Hifair™ II Reverse Transcriptase (Cat#11110ES) and various DNA Polymerases. Compared to human-derived RNase inhibitors, this product lacks two cysteine residues found in the human protein that are highly sensitive to oxidation, thus possessing higher antioxidant activity. Meanwhile, it is more suitable for experiments sensitive to high DTT concentrations, such as qPCR.
This product can be used in experiments such as first-strand cDNA synthesis, polysome separation, in vitro transcription, and cell-free translation systems.
Specifications
|
Cat.No. |
10610ES01 / 10610ES03 / 10610ES10 / 10610ES50 / 10610ES76 |
|
Size |
50 μL / 1 mL / 10 mL / 50 mL / 500 mL |
Components
|
Name |
10610ES01 （50 μL） |
10610ES03 （1 mL） |
10610ES10 （10 mL） |
10610ES50 (50 mL) |
10610ES76 (500 mL) |
|
Murine RNase Inhibitor (200 U/µL) |
50 μL |
1 mL |
10 mL |
50 mL |
10×50 mL |
Unit Definition
The amount of enzyme required to inhibit 50% of the activity of 5 ng RNase A is defined as 1 U. RNase A activity is determined quantitatively by the hydrolysis of Cyclic 2',3'-CMP to 3'-CMP.
Enzyme Storage Buffer Composition
20 mM Hepes-KOH, 150 mM KCl, 8 mM DTT, 0.5% NP40, 0.5% Tween-20, 50% Glycerol, pH 7.5 ± 0.2 25°C.
Storage Conditions
Store at -25~-15°C, valid for 2 years.
Precautions
1. This product inhibits RNase activity over a broad pH range, exhibiting maximum activity at pH 7-8.
2. Avoid foaming or vigorous stirring, vortexing, etc., to prevent inactivation of this product.
3. This product does not inhibit RNase H activity.
4. For your safety and health, please wear a lab coat and disposable gloves during operation.
Application Example (Reverse Transcription Experiment)
1. Prepare the following mixture in an RNase-free centrifuge tube:
|
Component |
Volume (μL) |
|
RNase-free ddH₂O |
to 20 |
|
5×Hifair™ II Buffer |
4 |
|
Oligo(dT)18 (50 μM) |
1 |
|
dNTP Mix (10 mM each) |
1 |
|
Murine RNase Inhibitor (200 U/μL) |
0.2 |
|
Hifair™ II Reverse Transcriptase (200 U/μL) |
1 |
|
Template RNA |
Total RNA: 1 ng-5 μg |
2. Mix gently;
3. 42°C 45 min, 85°C 5 min;
4. Store the product at -20°C.
Documents:
Manuals:
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