Recombinant Cynomolgus/Rhesus Macaque CD38 Protein, CF Summary
Details of Functionality |
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 >2,900 pmol/min/μg, as measured under the described conditions. |
Source |
Human embryonic kidney cell, HEK293-derived cynomolgus monkey CD38 protein Leu44-Ile301, with a C-terminal 6-His tag |
Accession # |
|
N-terminal Sequence |
Leu44 |
Protein/Peptide Type |
Recombinant Enzymes |
Purity |
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining. |
Endotoxin Note |
<0.10 EU per 1 μg of the protein by the LAL method. |
Applications/Dilutions
Dilutions |
|
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 |
36-44 kDa, reducing conditions |
Packaging, Storage & Formulations
Storage |
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 MES and NaCl. |
Purity |
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining. |
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). Per Well: - rcynoCD38: 0.1 µg
- Substrate: 200 µM
|
Notes
This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.
Alternate Names for Recombinant Cynomolgus/Rhesus Macaque CD38 Protein, CF
Background
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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