Measured by its ability to convert the substrate nicotinamide guanine dinucleotide (NGD+) to cyclic GDP-ribose. Graeff, R.M. et al. (1994) J. Biol. Chem. 269:30260. The specific activity is >20 pmol/min/µg, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived human CD157 protein Gly29-Lys292, with a C-terminal 6-His tag
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.
Applications/Dilutions
Dilutions
Enzyme Activity
Theoretical MW
31 kDa. Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
SDS-PAGE
38 kDa and 40 kDa, reducing conditions
Publications
Read Publications using 4736-AC in the following applications:
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
6 months from date of receipt, -20 to -70 °C as supplied.
3 months, -20 to -70 °C under sterile conditions after opening.
Buffer
Supplied as a 0.2 μm filtered solution in Tris and NaCl.
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Assay Procedure
Assay Buffer: 50 mM MES, 2.5 mM ZnCl2, pH 6.5
Recombinant Human CD157 (rhCD157) (Catalog # 4736-AC)
Substrate: Nicotinamide guanine dinucleotide (Sigma, Catalog # N-5131), 10 mM stock in deionized water
F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
Dilute rhCD157 to 5 µg/mL in Assay Buffer.
Dilute Substrate to 200 µM in Assay Buffer.
Load into a black well plate 50 µL of 5 µg/mL rhCD157, and start the reaction by adding 50 µL of 200 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL Substrate.
Incubate for 60 minutes at room temperature.
Read at excitation and emission wavelengths of 300 nm and 410 nm (top read), respectively, in endpoint mode.
Calculate specific activity:
Specific Activity (pmol/min/µg) =
Adjusted Fluorescence* (RFU) x Conversion Factor** (pmol/RFU)
Incubation time (min) x amount of enzyme (µg)
*Adjusted for Substrate Blank **Derived using calibration standard cyclic GDP ribose.
Per Well:
rhCD157: 0.25 µg
Substrate: 100 µM
Notes
This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.
Alternate Names for Recombinant Human CD157 Protein, CF
ADP-ribosyl cyclase 2
Bone marrow stromal antigen 1
bone marrow stromal cell antigen 1
BP-3
BST1
BST-1
cADPr hydrolase 2
CD157 antigen
CD157
Cyclic ADP-ribose hydrolase 2
EC 3.2.2.5
NAD(+) nucleosidase
Background
CD157, also known as bone marrow stromal cell antigen 1 (BST-1), is a glycosyl phosphatidylinositol anchored membrane protein that belongs to the CD38 family (1). CD157 was discovered in a bone marrow stromal cell line where it facilitates pre-B-cell growth (2, 3). Along with CD38, CD157 is a bifunctional ectoenzyme that exhibits both ADP-ribosyl cyclase and cyclic ADP ribose hydrolase activities (2). It may play a role in rheumatoid arthritis (RA) due to its enhanced expression in RA‑derived bone marrow stromal cell lines (3). CD157 has been predicted to function as a cell surface receptor and an immunoregulatory molecule (4).
Hussain, A. M. M. et al. (1998) Protein Express. Purif. 12:133.
Sato, A. et al. (1999) Biochem. J. 337:491.
Kaisho, T. et al. (1994) Proc. Natl. Acad. Sci. USA 91:5325.
Ortolan, E. et al. (2002) Cell Biochem. Funct. 20:309.
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