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Recombinant Human Adenosine Deaminase 2/CECR1 Protein, CF

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Summary
Reactivity HuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

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Recombinant Human Adenosine Deaminase 2/CECR1 Protein, CF Summary

Details of Functionality
Measured by the ability to catalyze the hydrolytic deamination of adenosine to inosine. The specific activity is >14,000 pmol/min/μg, as measured under the described conditions.
Source
Chinese Hamster Ovary cell line, CHO-derived human Adenosine Deaminase 2/CECR1 protein
Ile30-Lys511, with a C-terminal 6-His tag
Accession #
N-terminal Sequence
Ile30
Protein/Peptide Type
Recombinant Enzymes
Gene
CECR1
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
57 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
58-66 kDa, reducing conditions
Publications
Read Publications using
7518-AD in the following applications:

Packaging, Storage & Formulations

Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.
Buffer
Supplied as a 0.2 μm filtered solution in HEPES, NaCl, Glycerol.
Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Assay Procedure
  • 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
  1. Dilute rhCECR1 to 0.5 µg/mL in Assay Buffer.
  2. Dilute Substrate to 2 mM in Assay Buffer.
  3. In plate, combine 50 µL dilute rhCECR1 with 50 µL dilute substrate. Include a substrate control containing 50 µL dilute enzyme only.
  4. Incubate reactions at room temperature for 10 minutes.
  5. Add 100 µL stop/detection reagent to all wells used. Add 50 µL substrate to substrate control wells.
  6. Incubate for 30 minutes at room temperature in the dark to fully develop.
  7. Read with excitation and emission wavelengths of 330 nm and 450 nm (top read), respectively, in endpoint mode.
  8. 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).

Per Reaction:
  • rhCECR1: 0.025 µg
  • Adenosine: 1 mM

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human Adenosine Deaminase 2/CECR1 Protein, CF

  • ADA2
  • Adenosine Deaminase 2
  • adenosine deaminase CECR1
  • ADGF
  • ADGFadenosine deaminase 2
  • cat eye syndrome chromosome region, candidate 1
  • Cat eye syndrome critical region protein 1
  • CECR1
  • CECR2
  • EC 3.5.4.4
  • IDGFL

Background

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).
  1. Wolfenden, R.V. et al. (1969) Biochemistry 6:2412.
  2. Lowenstein, J.M. (1972) Physiol. Rev. 52:382.
  3. Riazi, M.A. et al. (2000) Genomics 64:277.
  4. Zavialov, A.V. and A. Engstrom (2005) Biochem. J. 391:51.
  5. Zavialov, A.V. et al. (2010) J. Leucoc. Biol. 88:279.

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Recombinant Human
Adenosine Deaminase 2 ...
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Recombinant Human Adenosine Deaminase 2/CECR1 Protein, CF
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Publications for Adenosine Deaminase 2/CECR1 (7518-AD)(3)

We have publications tested in 3 confirmed species: Human, Mouse, N/A.

We have publications tested in 1 application: Bioassay.


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(3)
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(1)
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(1)
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Bioinformatics

Gene Symbol CECR1
Uniprot