Kell, a type II membrane glycoprotein, is linked through a single disulfide bond to XK, a putative membrane transporter. The two proteins constitute the Kell blood group antigens (1). Kell is a zinc endopeptidase of the neprilysin (NEP) family which also includes endothelin converting enzymes (ECE-1 and ECE-2), PEX, XCE, DINE and several NEP-like proteins (2). It has been shown to cleave big endothelin-3 (ET-3) at Trp21-Ile22, yielding ET-3, and to a much lesser extent, big ET-1 and big ET-2 at Trp21-Val22, yielding ET-1 and ET-2 (3). Several different molecular defects cause the Kell null phenotype, which has no obvious clinical outcome (4).
Recombinant Mouse Kell Protein, CF
R&D Systems | Catalog # 1454-ZN
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Key Product Details
- R&D Systems NS0-derived Recombinant Mouse Kell Protein (1454-ZN)
- 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
Applications
Enzyme Activity
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Product Specifications
Source
Mouse myeloma cell line, NS0-derived mouse Kell protein
Phe50-Trp713, with an N-terminal 7-His tag
Phe50-Trp713, with an N-terminal 7-His tag
Purity
>95%, 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
76 kDa
SDS-PAGE
106 kDa, reducing conditions
Activity
Measured by its ability to cleave the fluorogenic peptide substrate, Mca-RPPGFSAFK(Dnp)-OH (Catalog # ES005).
The specific activity is >0.2 pmol/min/µg, as measured under the described conditions.
Formulation, Preparation, and Storage
1454-ZN
| Formulation | Supplied as a 0.2 μm filtered solution in Tris, NaCl and ZnCl2. |
| 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: Kell
References
- Lee, S. et al. (2000) Transfus. Med. Rev. 14:93.
- Turner, A.J. et al. (2001) BioEssays. 23:261.
- Lee, S. et al. (1999) Blood 94:1440.
- Lee, S. et al. (2001) J. Biol. Chem. 276:27281.
Long Name
Zinc protease blood group antigen
Alternate Names
CD238, KEL
Gene Symbol
KEL
UniProt
Additional Kell Products
Product Documents for Recombinant Mouse Kell 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 Kell Protein, CF
For research use only
Related Research Areas
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Protocols
View specific protocols for Recombinant Mouse Kell Protein, CF (1454-ZN):
Materials
- Assay Buffer: 0.1 M MES, pH 6.5
- Recombinant Mouse Kell (rmKell) (Catalog # 1454-ZN)
- Substrate: MCA-Arg-Pro-Pro-Gly-Phe-Ser-Ala-Phe-Lys(DNP)-OH (Catalog # ES005), 2 mM stock in DMSO
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rmKell to 20 ng/µL in Assay Buffer.
- Dilute Substrate to 20 µM in Assay Buffer.
- Load 50 µL of the 20 ng/µL rmKell in a plate, and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 20 µM Substrate.
- Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively, in kinetic mode for 1 hour.
- Calculate specific activity:
|
Specific Activity (pmol/min/µg) = |
Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU) |
| amount of enzyme (µg) |
*Adjusted for Substrate Blank
**Derived using calibration standard MCA-Pro-Leu-OH (Bachem, Catalog # M-1975).
Per Well:
- rmKell: 1 µg
- Substrate: 10 µM