Recombinant Human Adenosine Deaminase 2/CECR1 Protein, CF Summary
Ile30-Lys511, with a C-terminal 6-His tag
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 HEPES, NaCl, Glycerol.|
|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: 25 mM Sodium phosphate, 1 M NaCl, pH 6.0
- Recombinant Human Adenosine Deaminase 2/CECR1 (rhCECR1) (Catalog # 7518-AD)
- Substrate: Adenosine (Sigma, Catalog # A9251), 10 mM stock in deionized water (incubate 10 minutes at 37 °C to fully solubilize)
- Stop/detection reagent: 0.2 M Sodium Hydroxide, 15 mM ortho-phthaldehyde (Sigma, Catalog # P0657), 0.1% beta -mercaptoethanol
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rhCECR1 to 0.5 µg/mL in Assay Buffer.
- Dilute Substrate to 2 mM in Assay Buffer.
- In plate, combine 50 µL dilute rhCECR1 with 50 µL dilute substrate. Include a substrate control containing 50 µL dilute enzyme only.
- Incubate reactions at room temperature for 10 minutes.
- Add 100 µL stop/detection reagent to all wells used. Add 50 µL substrate to substrate control wells.
- Incubate for 30 minutes at room temperature in the dark to fully develop.
- Read with excitation and emission wavelengths of 330 nm and 450 nm (top read), respectively, in endpoint mode.
- Specific activity may be determined using the following equation:
Specific Activity (pmol/min/µg) =
Adjusted Fluorescence* (RFU) x Conversion Factor** (pmol/RFU) Incubation time (min) x amount of enzyme (µg)
*Adjusted for substrate control
**Derived using calibration standard ammonium sulfate (Amresco, Catalog # 0191).
- rhCECR1: 0.025 µg
- Adenosine: 1 mM
Background: Adenosine Deaminase 2/CECR1
Adenosine deaminase is one of the key enzymes of purine nucleotide metabolism. It catalyses the conversion of adenosine and deoxy-adenosine to inosine and deoxy-inosine, respectively (1, 2). Adenosine Deaminase 2 (ADA2) is also known as CECR1 because it is a candidate gene for cat eye syndrome, a developmental disorder (3). ADA is a secreted protein that is expressed in many tissues, with the highest expression in lymphoblasts, heart, lung, and placenta (4). ADA2 is a member of a family of adenosine deaminase-related growth factors (ADGFs), proteins that are involved in tissue development (4). ADA2 induces the differentiation of monocytes into macrophages and stimulates the proliferation of T helper cells and macrophages by a mechanism independent of its catalytic activity (5). It has been suggested that ADA2 could be a therapeutic target for the control of immune responses in inflammation and cancer (5).
- Wolfenden, R.V. et al. (1969) Biochemistry 6:2412.
- Lowenstein, J.M. (1972) Physiol. Rev. 52:382.
- Riazi, M.A. et al. (2000) Genomics 64:277.
- Zavialov, A.V. and A. Engstrom (2005) Biochem. J. 391:51.
- Zavialov, A.V. et al. (2010) J. Leucoc. Biol. 88:279.
Citations for Recombinant Human Adenosine Deaminase 2/CECR1 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.
Citations: Showing 1 - 2
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Cellular sensing of extracellular purine nucleosides triggers an innate IFN-&beta response
Authors: R Dhanwani, M Takahashi, IT Mathews, C Lenzi, A Romanov, JD Watrous, B Pieters, CC Hedrick, CA Benedict, J Linden, R Nilsson, M Jain, S Sharma
Sci Adv, 2020-07-22;6(30):eaba3688.
Sample Types: Whole Cells
Adenosine deaminase-1 delineates human follicular helper T cell function and is altered with HIV
Authors: V Tardif, R Muir, R Cubas, M Chakhtoura, P Wilkinson, T Metcalf, R Herro, EK Haddad
Nat Commun, 2019-02-18;10(1):823.
Sample Types: Whole Cells
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