Recombinant Human Carnosine Dipeptidase 1 Protein, CF Summary
Pro28-His507, with a C-terminal 10-His tag
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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 Tris and NaCl.|
|Shipping||The product is shipped with dry ice or equivalent. 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 Tris, pH 7.5
- Recombinant Human Carnosine Dipeptidase 1/CNDP1 (rhCPGL2) (Catalog # 2489-ZN)
- Substrate: L-Carnosine (Sigma, Catalog # C9625), 100 mM stock in deionized water
- Trichloroacetic acid (TCA) (Sigma, Catalog # T6399), 10% in deionized water
- NaOH, 2 M stock in deionized water
- o-Phthaldialdehyde (OPA) (Sigma, Catalog # P0657), 50 mg/mL in DMSO
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rhCDNP1 to 2 ng/µL in Assay Buffer.
- Dilute Substrate to 2 mM in Assay Buffer.
- Mix 50 μL of 2 ng/µL rhCDNP1 and 50 μL 2 mM Substrate for a final concentration of 1 ng/µL and 1 mM respectively. Include a Substrate Blank containing 50 μL rhCDNP1, 50 μL 1% TCA added 1 minute prior to substrate addition, and 50 μL Substrate.
- Incubate for 1 hour at room temperature.
- Stop reaction by adding 50 μL 1% TCA (except the Substrate Blank). Centrifuge for 10 minutes at 13,000 rpm in a microcentrifuge to spin down any precipitate.
- Transfer the supernatant to new tubes.
- Dilute OPA to 5 mg/mL in 2 M NaOH.
- Add 50 μL of 5 mg/mL OPA to each reaction.
- Incubate for 30 minutes at room temperature.
- Load 200 µL (entire volume) of the incubated samples into the plate.
- Read at excitation and emission wavelengths of 360 nm and 460 nm (top read), respectively, in endpoint mode.
- 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 L-Histidine (Sigma, Catalog # H6034).
- rhCDNP1: 0.100 µg
- Substrate: 0.5 mM
Background: Carnosine Dipeptidase 1/CNDP1
The human CNDP1 gene encodes carnosine dipeptidase 1, a member of the M20 family of metalloproteases (1, 2). Also known as X-His dipeptidase, glutamate carboxypeptidase-like protein 2 (CPGL-2) or carnosinase 1 (CN1), CNDP1 is a secreted dipeptidase with a narrow specificity for Xaa-His dipeptides including those with Xaa = beta -Ala (carnosine) and Xaa = gamma -aminobutyric acid (homocarnosine), two naturally occurring dipeptides with potential neuroprotective and neurotransmitter fucntions in the brain. In comparison, a closely related protein known as CNDP2, CPGL or CN2, is a cytosolic nonspecific dipeptidase.
- Teufel, M. et al. (2004) J. Biol. Chem. 278:6521.
- Bauer, K. (2004) in Handbook of Proteolytic Enzymes (ed. Barrett, et al.) p. 1022, Academic Press, San Diego.
Citation for Recombinant Human Carnosine Dipeptidase 1 Protein, CF
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
1 Citation: Showing 1 - 1
Identification and validation of novel cerebrospinal fluid biomarkers for staging early Alzheimer's disease.
Authors: Perrin RJ, Craig-Schapiro R, Malone JP, Shah AR, Gilmore P, Davis AE, Roe CM, Peskind ER, Li G, Galasko DR, Clark CM, Quinn JF, Kaye JA, Morris JC, Holtzman DM, Townsend RR, Fagan AM
PLoS ONE, 2011;6(1):e16032.
Sample Types: N/A
Applications: ELISA (Standard)
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