p19 siRNA Binding ProteinProduct information
p19 siRNA Binding Protein
|1.000 units ( 10000 units/ml )||-||Unavailable in your region|
- For the isolation of siRNA
- Isolated from a recombinant source
Product Sourcep19 siRNA Binding Protein is cloned and expressed in E. coli as a fusion protein with an amino terminal MBP (maltose binding protein) and a carboxy terminal CBD (chitin binding domain).
The following reagents are supplied with this product:
|Store at (°C)||Concentration|
|p19 siRNA Binding Buffer (200 μl)||2X|
Advantages and Features
ApplicationsHigh affinity binding of siRNAs
Affinity purification of siRNA with chitin magnetic beads
Properties and Usage
Unit DefinitionOne unit is defined as the amount of protein that binds to 10 ng of siRNA at 25°C in 1 hour.
Unit Assay Conditions
1 unit of p19 siRNA Binding Protein was incubated with 20 ng of phosphorylated siRNA in 1X p19 siRNA Binding Buffer in a volume of 20 µl at 25°C for 1 hour.
10 mM Tris-HCl
150 mM NaCl
0.5 mM DTT
1 mM EDTA
pH 7.5 @ 25°C
Quality Assurance Statement
- p19 siRNA Binding Protein contains no detectable DNases, RNases and phosphatases. The purified protein contains no detectable DNA or RNA as determined by ethidium bromide staining of an agarose gel.
Quality Control AssaysThe following Quality Control Tests are performed on each new lot and meet the specifications designated for the product. Individual lot data can be found on the Product Summary Sheet/Datacard or Manual which can be found in the Supporting Documents section of this page. Further information regarding NEB product quality can be found here.
- Endonuclease Activity (Nicking):
The product is tested in a reaction containing a supercoiled DNA substrate. After incubation for 4 hours the percent converted to the nicked form is determined by agarose gel electrophoresis.
- Exonuclease Activity (Radioactivity Release):
The product is tested in a reaction containing a radiolabeled mixture of single and double-stranded DNA. After incubation for 4 hours the exonuclease activity is determined by the % release of radioactive nucleotides.
- Non-Specific DNase Activity (16 hour):
The product is tested for non-specific nuclease degradation in a reaction containing a DNA substrate. After incubation for 16 hours there is no detectable degradation of the DNA substrate as determined by agarose gel electrophoresis.
- This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
Research Use Only
- The buyer/user has a non-exclusive license to use the vector for Research Purposes Only. Commercial use of this vector requires a license from New England Biolabs, Inc.
U.S. Patent No. 5,643,758
Appln. No. WO2007/120809
While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.
For more information about commercial rights, please contact NEB's Global Business Development team at email@example.com.
This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
TrademarksNEW ENGLAND BIOLABS® is a registered trademark of New England Biolabs, Inc.
p19 siRNA Binding Protein
p19 siRNA Binding Buffer (200 μl)
- p19 siRNA Binding Protein can selectively bind siRNAs in the presence of a 2,000 fold excess of other RNAs.
- 1X p19 siRNA Binding Buffer:
20 mM Tris-HCl, 100 mM NaCl, 1 mM EDTA, 1 mM TCEP, 0.02% Tween-20 (pH 7.0 @ 25°C)
- p19 siRNA Binding Protein does not bind ssRNA.Double-stranded 21 bp RNA with a 2-base overhang and a 5´ phosphate binds the most efficiently. Double-stranded miRNAs with mismatched base pairs will bind with lower affinity.
- Due to the difference in molecular weight between p19 siRNA Binding Protein and siRNA, a 10 to 20-fold excess of p19 siRNA Binding Protein is needed.
- TCEP (Tris 2-carboxyethyl Phosphine) can be replaced with DTT (dithiotreitol) in the required solutions. However, if DTT is used, it needs to be added separately into the reaction.
- Silhavy, D. et al. (2002). EMBO J. 21
- Vargason, J.M., et al. (2003). Cell. 115
- Qiu, W. et al. (2002). Mol. Plant Microbe Interact. 15