Recombinant Mouse CD157 Protein, CF

• Reactivity: Mu
• Applications: Enzyme Activity
• Format: Carrier-Free

Recombinant Mouse CD157 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 >10 pmol/min/µg, as measured under the described conditions.
• Source: Mouse myeloma cell line, NS0-derived mouse CD157 protein
  Ala25-Glu285, with a C-terminal 6-His tag
• Accession #: Q64277
• N-terminal Sequence: Ala25
• Protein/Peptide Type: Recombinant Enzymes
• Gene: Bst1
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Endotoxin Note: <1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Enzyme Activity
• Theoretical MW: 30 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: 37 kDa, 40 kDa and 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 Tris 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, 2.5 mM ZnCl2, pH 6.5
  • Recombinant Mouse CD157 (rmCD157) (Catalog # 4710-AC)
  • Substrate: Nicotinamide guanine dinucleotide (NGD⁺) (Sigma, Catalog # N-5131), 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
  1. Dilute rmCD157 to 5 µg/mL in Assay Buffer.
  2. Dilute Substrate to 200 µM in Assay Buffer.
  3. Into plate load 50 µL of 5 µg/mL rmCD157 and start the reaction by adding 50 µL of 200 µM Substrate. Include a blank control containing 50 µL Assay Buffer and 50 µL Substrate.
  4. Mix well.
  5. Incubate for 90 minutes at room temperature.
  6. Read at excitation and emission wavelengths of 300 nm and 410 nm (top read), respectively in endpoint mode.
  7. Calculate specific activity:

  Specific Activity (pmol/min/µg) = (Adjusted Fluorescence* (RFU) x Conversion Factor** (pmol/RFU)) / (Incubation time (min) x amount of enzyme (µg))

  *Adjusted for Substrate Blank

  **Derived using calibration standard standard cyclic GDP ribose. Per Well:

  • rmCD157: 0.25 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 Mouse CD157 Protein, CF

• ADP-ribosyl cyclase 2
• Bone marrow stromal antigen 1
• bone marrow stromal cell antigen 1
• BP-3
• BST1
• BST-1
• cADPr hydrolase 2
• CD157 antigen
• CD157
• Cyclic ADP-ribose hydrolase 2
• EC 3.2.2.5
• NAD(+) nucleosidase

Background

CD157, also known as bone marrow stromal cell antigen 1 (BST-1), is a glycosyl phosphatidylinositol anchored membrane protein that belongs to the CD38 family (1). CD157 was discovered in a bone marrow stromal cell line where it facilitates pre-B-cell growth (2, 3). Along with CD38, CD157 is a bifunctional ectoenzyme that exhibits both ADP-ribosyl cyclase and cyclic ADP ribose hydrolase activities (2). It may play a role in rheumatoid arthritis (RA) due to its enhanced expression in RA‑derived bone marrow stromal cell lines (3). CD157 has been predicted to function as a cell surface receptor and an immunoregulatory molecule (4).

1. Hussain, A. M. M. et al. (1998) Protein Express. Purif. 12:133.
2. Sato, A. et al. (1999) Biochem. J. 337:491.
3. Kaisho, T. et al. (1994) Proc. Natl. Acad. Sci. USA 91:5325.
4. Ortolan, E. et al. (2002) Cell Biochem. and Function. 20:309.

Bioinformatics

• Gene Symbol: Bst1
• Uniprot:
  • Q64277(Mouse)