Recombinant Cynomolgus/Rhesus Macaque CD38 Protein, CF Summary
Product Specifications
Leu44-Ile301, 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.
9834-AC
Formulation | Supplied as a 0.2 μm filtered solution in MES 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.
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Assay Procedure
- Assay Buffer: 50 mM MES, pH 6.5
- Recombinant (cynoCD38) (Catalog # 9834-AC)
- Substrate: Nicotinamide guanine dinucleotide sodium salt (NGD+) (Sigma, Catalog # N5131), 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 rcynoCD38 to 2 ng/µL in Assay Buffer.
- Dilute Substrate to 400 µM in Assay Buffer.
- Load in plate 50 µL of 2 ng/µL rcynoCD38, and start the reaction by adding 50 µL of 400 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 400 µM Substrate.
- Read at excitation and emission wavelengths of 300 nm and 410 nm (top read), respectively, in kinetic mode of 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).
- rcynoCD38: 0.1 µg
- Substrate: 200 µM
Reconstitution Calculator
Background: CD38
CD38, also known as ADP‑ribosyl cyclase, converts NAD(P)+ into three separate products with calcium mobilizing ability: cyclic ADP-ribose, NAADP+, and ADP‑ribose (1). CD38 is a Type II transmembrane glycoprotein composed of an intracellular domain, a single transmembrane helix domain, and a large extracellular domain that contains the catalytic site (2). Within the extracellular domain, cynoCD38 shares 93% amino acid sequence identity with human CD38. CD38 is expressed in B and T lymphocytes, osteoclasts, and in cardiac, pancreatic, liver and kidney cells (3, 4). Through its production of cyclic ADP‑ribose, CD38 modulates calcium‑mediated signal transduction in many types of cells (5, 6). CD38 is also reported to bind as a receptor to trigger signaling cascades (7, 8). Through both mechanisms, CD38 influences proliferation and trafficking (8, 9). CD38 is used as a marker for poor prognosis in chronic lymphocytic leukemia and multiple myeloma and is an attractive cancer immunotherapy drug target (8-11).
- Schuber, F. and F.E. Lund (2004) Curr. Mol. Med. 4:249.
- Liu, Q. et al. (2005) Structure. 13:1331.
- 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.
- Deaglio, S. (2000) Chem. Immunol. 75:99.
- Chillemi, A. et al. (2013) Mol. Med. 19:99.
- Vaisitti, T. et al. (2015) Leukemia. 29:356.
- Atanackovic, D. et al. (2016) Oncoimmunology. 5:e1217374.
- Tai, Y-T. et al. (2017) Oncotarget. 8:112166.
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