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p19 siRNA Binding Protein
Catalog #
M0310S
Size
1,000 units
Concentration
10,000 units/ml
Price
$78.00
Qty
M0310L
5,000 units
10,000 units/ml
$312.00
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1
Categories: miRNA & siRNA Products, RNA Isolation & Detection Products
Applications: RNA Detection & Isolation
Product
Information
Protocols &
Manuals
Other Tools &
Resources
Description
Properties and Usage
Notes
Quality &
Safety
Legal
Information
Advantages and Features
Related Products
References
Description
The p19 siRNA Binding Protein (19 kDa) from the Carnation Italian Ringspot Virus (CIRV) plant binds siRNAs with nanomolar affinity(1). The
dimeric protein binds 21 base siRNAs with a 2 base 3´ extension and a 5´phosphate. The protein binds RNA in a size dependent and
sequence independent manner. If the siRNAs are 4 bases longer, the affinity for the protein is reduced about 100-fold (2). When p19 siRNA
Binding Protein is expressed in plants it suppresses RNA interference (3).
Figure 1:
Size specific binding of siRNA by p19 siRNA Binding Protein. Three phosphorylated dsRNAs of 17,21 and 25 bases (30 ng each band) were
mixed with increasing amount of p19 siRNA Binding Protein (0-3 µg) in a 20 µl reaction, and incubated at room temperature for 1 hour. The
reaction was analyzed on a 20% polyacrylamide gel stained with ethidium bromide. Marker M is the siRNA Marker (NEB #N2101 ).
Highlights
For the isolation of siRNA
Isolated from a recombinant source
Product Source
p19 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).
Reagents Supplied
The following reagents are supplied with this product:
Store at (°C)
p19 siRNA Binding Buffer (200 μl)
Concentration
2X
Advantages and Features
Applications
High affinity binding of siRNAs
Affinity purification of siRNA with chitin magnetic beads
Properties and Usage
Unit Definition
One 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.
Storage Temperature
-20°C
Storage Conditions
10 mM Tris-HCl
150 mM NaCl
0.5 mM DTT
1 mM EDTA
50% Glycerol
pH 7.5 @ 25°C
Related Products
Companion Products
12-Tube Magnetic Separation Rack
6-Tube Magnetic Separation Rack
Anti-CBD Monoclonal Antibody
Anti-MBP Monoclonal Antibody
Chitin Resin
Chitin Magnetic Beads
Notes
1. p19 siRNA Binding Protein can selectively bind siRNAs in the presence of a 2,000 fold excess of other RNAs.
2. 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)
3. 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.
4. 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.
5. 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.
References
1. Silhavy, D. et al. (2002). EMBO J. 21
2. Vargason, J.M., et al. (2003). Cell. 115
3. Qiu, W. et al. (2002). Mol. Plant Microbe Interact. 15
Protocols
Protocols
Datacards
1. Protocol for isolation of siRNA from p19 siRNA Binding Protein (M0310) using chitin magnetic beads
Datacards
The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and
quality controls. The following file naming structure is used to name the majority of these document files: [Catalog
Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at [email protected] or fill out the
Technical Support Form for appropriate document.
M0310Datasheet-Lot0031205
M0310Datasheet-Lot0031402
M0310Datasheet-Lot0031405
M0310Datasheet-Lot0031410
M0310Datasheet-Lot0031501
Selection Charts
Interactive Tools
Selection Charts
Reagents for RNA Library Preparation
Interactive Tools
NEBioCalculator
Quality Control
Datacards
Safety Data Sheet
Quality Control
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 Assays
The 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.
Safety Data Sheet
The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.
p19 siRNA Binding Protein
p19 siRNA Binding Buffer (200 μl)
Datacards
The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and
quality controls. The following file naming structure is used to name the majority of these document files: [Catalog
Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at [email protected] or fill out the
Technical Support Form for appropriate document.
M0310Datasheet-Lot0031205
M0310Datasheet-Lot0031402
M0310Datasheet-Lot0031405
M0310Datasheet-Lot0031410
M0310Datasheet-Lot0031501
Legal and Disclaimers
Legal and Disclaimers
This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc
(NEB).
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 protected].
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
Trademarks
NEW ENGLAND BIOLABS® is a registered trademark of New England Biolabs, Inc.
Disclaimers
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.