Recombinant Human CD38 His-tag Avi-tag Protein, CF Summary
Accession # P28907.2
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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 PBS.|
|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, pH 6.5
- Recombinant Human CD38 His-tag Avi-tag (rhCD38/His/Avi-tag) (Catalog # AVI2404)
- Substrate: Nicotinamide guanine dinucleotide (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 rhCD38/His/Avi-tag to 2 ng/µL in Assay Buffer.
- Dilute Substrate to 200 µM in Assay Buffer.
- Load in plate, 50 µL of 2 ng/µL rhCD38/His/Avi-tag, 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).
- rhCD38/His/Avi-tag: 0.1 µg
- Substrate: 100 µM
2 μg/lane of Recombinant Human CD38 His-tag Avi-tag Protein (AVI2404) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 40-52 kDa.
CD38, also known as ADP-ribosyl cyclase, 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). Our Avi-tag Biotinylated Recombinant Human CD38 features biotinylation at a single site contained within the Avi-tag, a unique 15 amino acid peptide. Protein orientation will be uniform when bound to streptavidin-coated surface due to the precise control of biotinylation and the rest of the protein is unchanged so there is no interference in the protein's bioactivity.
- 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.
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