Recombinant Human CD38 Fc Chimera 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 >900 pmol/min/μg, as measured under the described conditions. |
Source |
Human embryonic kidney cell, HEK293-derived human CD38 protein Human CD38 (Val43-Ile300) Accession # P28907.2 | IEGRMD | Human IgG1 (Pro100-Lys330) | N-terminus | | C-terminus | |
|
Accession # |
|
N-terminal Sequence |
Val43 |
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 |
57 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 |
67-74 kDa, under 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 Human CD38 (rhCD38) (Catalog # 10920-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 M5 by Molecular Devices) or equivalent
- Dilute rhCD38 to 4 ng/µL in Assay Buffer.
- Dilute Substrate to 400 µM in Assay Buffer.
- Load in plate 50 µL of 4 ng/µL rhCD38, 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 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).
Per Well: - rhCD38: 0.2 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 Human CD38 Fc Chimera 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). 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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