Serpin A9, also known as Centerin, is a member of the Serpin superfamily of serine protease inhibitors (1). Serpins are the most broadly distributed superfamily of protease inhibitors. They are known to inhibit serine proteases and some are known to inhibit caspases and papain-like cysteine proteases (1). Recombinant mouse Serpin A9 shows inhibition towards trypsin, thrombin, and plasmin and binds DNA and heparin (2). Its expression is limited to germinal center B cells and lymphoid malignancies with germinal center B-cell maturation (3). New findings suggest that Serpin A9 could be used as a potential diagnostic marker in the recognition of various germinal center derived lymphomas (4).
Recombinant Mouse Serpin A9/Centerin Protein, CF
R&D Systems | Catalog # 5679-PI
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Key Product Details
- R&D Systems NS0-derived Recombinant Mouse Serpin A9/Centerin Protein (5679-PI)
- Quality control testing to verify active proteins with lot specific assays by in-house scientists
- All R&D Systems proteins are covered with a 100% guarantee
Source
NS0
Accession Number
Structure / Form
Monomer
Applications
Inhibition Activity
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Product Specifications
Source
Mouse myeloma cell line, NS0-derived mouse Serpin A9/Centerin protein
Asn26-Met418, with an N-terminal 6-His tag
Accession # Q9D7D2
Asn26-Met418, with an N-terminal 6-His tag
Accession # Q9D7D2
Purity
>90%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
N-terminal Sequence Analysis
His
Predicted Molecular Mass
45 kDa
SDS-PAGE
40-50 kDa, reducing conditions
Activity
Measured by its ability to inhibit trypsin cleavage of a fluorogenic peptide substrate, Mca-RPKPVE-Nval-WRK(Dnp)-NH2 (Catalog # ES002).
The IC50 value is <7 nM, measured under the described conditions.
The IC50 value is <7 nM, measured under the described conditions.
Formulation, Preparation, and Storage
5679-PI
| Formulation | Supplied as a 0.2 μm filtered solution in Tris and NaCl. |
| Shipping | The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
|
Background: Serpin A9/Centerin
References
Long Name
Serpin Peptidase Inhibitor, Clade A, Member 9
Alternate Names
Centerin, GCET1, SERPINA11b
Gene Symbol
SERPINA9
UniProt
Additional Serpin A9/Centerin Products
Product Documents for Recombinant Mouse Serpin A9/Centerin Protein, CF
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Recombinant Mouse Serpin A9/Centerin Protein, CF
For research use only
Related Research Areas
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Protocols
View specific protocols for Recombinant Mouse Serpin A9/Centerin Protein, CF (5679-PI):
Materials
- Assay Buffer: 50 mM Tris, 10 mM CaCl2, 0.15 M NaCl, 0.05% (v/v) Brij-35, pH 7.5 (TCNB)
- Recombinant Mouse Serpin A9/Centerin (rmSerpin A9) (Catalog # 5679-PI)
- Recombinant Human Active Trypsin 3/PRSS3 (rhTrypsin 3) (Catalog # 3714-SE)
- Fluorogenic Peptide Substrate: MCA-Arg-Pro-Lys-Pro-Val-Glu-NVAL-Trp-Arg-Lys(DNP)-NH2 (Catalog # ES002)
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rhTrypsin 3 to 1 ng/µL in Assay Buffer.
- Prepare a curve of rmSerpin A9 (MW: 44,977 Da) in Assay Buffer. Make the following serial dilutions: 1200 nM, 600 nM, 300 nM, 150 nM, 75 nM, 37.5 nM, 18.8 nM, 9.4 nM, 4.7 nM, and 2.3 nM.
- Combine equal volumes of dilute rhTrypsin 3 and rmSerpin A9 at each concentration of the curve. Include two controls containing equal volumes of Assay Buffer and diluted rhTrypsin 3 without any rmSerpin A9.
- Incubate mixtures at 37 °C for 30 minutes.
- Dilute the reaction mixtures 5-fold with Assay Buffer.
- Dilute Substrate to 20 µM with Assay Buffer.
- Load in a plate 50 µL of the incubated mixtures, and start the reaction by adding 50 µL of 20 mM Substrate.
- Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively, in kinetic mode for 5 minutes.
- Derive the 50% inhibition concentration (IC50) for rmSerpin A9 by plotting RFU/min (or specific activity) vs concentration with 4-PL fitting.
- Calculate specific activity of Trypsin at each point using the following formula (if needed):
|
Specific Activity (pmoles/min/µg) = |
Adjusted Vmax* (RFU/min) x Conversion Factor** (pmole/RFU) |
| amount of enzyme (µg) |
*Adjusted for Substrate Blank
**Derived using calibration standard MCA-Pro-Leu-OH (Bachem, Catalog # M-1975).
Per Well:
- rhTrypsin 3: 0.005 µg (2.1 nM)
- rmSerpin A9 curve: 60 nM, 30 nM, 15 nM, 7.5 nM, 3.8 nM, 1.9 nM, 0.94 nM, 0.47 nM, 0.23 nM, and 0.12 nM
- Substrate: 10 µM
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