Recombinant Mouse CD157 Protein, CF Summary
Product Specifications
Ala25-Glu285, with a C-terminal 6-His tag
Analysis
Product Datasheets
Carrier Free
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
4710-AC
| Formulation | Supplied as a 0.2 μm filtered solution in Tris and NaCl. |
| Shipping | The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage: | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
|
Assay Procedure
- Assay Buffer: 50 mM MES, 2.5 mM ZnCl2, pH 6.5
- Recombinant Mouse CD157 (rmCD157) (Catalog # 4710-AC)
- Substrate: Nicotinamide guanine dinucleotide (NGD+) (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 rmCD157 to 5 µg/mL in Assay Buffer.
- Dilute Substrate to 200 µM in Assay Buffer.
- Into plate load 50 µL of 5 µg/mL rmCD157 and start the reaction by adding 50 µL of 200 µM Substrate. Include a blank control containing 50 µL Assay Buffer and 50 µL Substrate.
- Mix well.
- Incubate for 90 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 standard cyclic GDP ribose. Per Well:
- rmCD157: 0.25 µg
- Substrate: 100 µM
Reconstitution Calculator
Background: CD157
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. and Function. 20:309.
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