Recombinant Human CD38 His-tag Avi-tag Protein, CF Summary
Additional Information |
Biotinylated |
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 >1500 pmol/min/μg, as measured under the described conditions. Measured by its binding ability in a functional ELISA. When Human CD38 Antibody
(Catalog #
AF2404) is immobilized at 1 µg/mL (100 µL/well), it binds to Biotinylated
Recombinant Human CD38 His-tag Avi-tag (Catalog # AVI2404) with an ED 50 of 1-6 ng/mL. |
Source |
Human embryonic kidney cell, HEK293-derived human CD38 protein Human CD38 (Val43-Ile300) Accession # P28907.2 | HHHHHH | Avi-tag | N-terminus | | C-terminus | |
|
N-terminal Sequence |
Val43 |
Structure / Form |
Biotinylated via Avi-tag |
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 |
- Binding Activity2
- Enzyme Activity
|
Theoretical MW |
33 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 |
40-52 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 PBS. |
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 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). Per Well: - rhCD38/His/Avi-tag: 0.1 µg
- Substrate: 100 µM
|
Notes
This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.
Alternate Names for Recombinant Human CD38 His-tag Avi-tag Protein, CF
Background
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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