Recombinant Mouse CD38 Protein, CF Summary
Leu45-Thr304, with a C-terminal 6-His tag
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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.
|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 Buffer: 50 mM MES, 50 mM NaCl, pH 6.5
- Recombinant Mouse CD38 (rmCD38) (Catalog # 4947-AC)
- Substrate: 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 rmCD38 to 0.1 ng/µL in Assay Buffer.
- Dilute Substrate to 200 µM in Assay Buffer.
- Load into a plate 50 µL of 0.1 ng/µL rmCD38, and start the reaction by adding 50 µL of 200 µM Substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of 200 µM Substrate.
- Read at excitation and emission wavelengths of 300 nm and 410 nm (top read), respectively, in kinetic mode for 5 minutes.
- 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 cyclic GDP ribose (cGDPR).
- rmCD38: 0.005 µg
- Substrate: 100 µM
CD38, also known as ADP-ribosyl cyclase and cyclic ADP-ribose hydrolase, is a Type II integral membrane protein. The enzyme is able to transform NAD(P)+ into three different products with calcium mobilizing ability; cyclic ADP-ribose, NAADP+, and ADP-ribose (1). CD38 is expressed in B and T lymphocytes, osteoclasts, and in cardiac, pancreatic, liver and kidney cells (2, 3). Through its production of cyclic ADP-ribose, CD38 modulates calcium-mediated signal transduction in many types of cells, including neutrophils and pancreatic beta cells (4, 5). CD38 has been shown to regulate oxytocin secretion, and may be involved in the development of complex social behaviors in mammals (6).
- Schuber, F. and F.E. Lund (2004) Curr. Mol. Med. 4:249.
- Jackson, D.G. and J.I. Bell (1990) J. Immunol. 144:2811.
- Sun, L. et al. (1999) J. Cell Biol. 146:1161.
- Partida-Sanchez, S. et al. (2001) Nature Med. 7:1209.
- Kato, I. et al. (1995) J. Biol. Chem. 270:30045.
- Jin, D. et al. (2007) Nature 446:41.
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