Recombinant Human Adenosine Deaminase/ADA Protein, CF

Catalog # Availability Size / Price Qty
7048-AD-010
Product Details
Citations (1)
FAQs
Reviews

Recombinant Human Adenosine Deaminase/ADA Protein, CF Summary

Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
Activity
Measured by the ability to catalyze the hydrolytic deamination of adenosine to inosine. The specific activity is >44,000 pmol/min/μg, as measured under the described conditions.
Source
Spodoptera frugiperda, Sf 21 (baculovirus)-derived human Adenosine Deaminase/ADA protein
Met1-Leu363, with a C-terminal 6-His tag
Accession #
N-terminal Sequence
Analysis
Inconclusive result, Met predicted. Protein identity confirmed by MS analysis of tryptic fragments
Predicted Molecular Mass
42 kDa
SDS-PAGE
40-42 kDa, reducing conditions

Product Datasheets

Carrier Free

What does CF mean?

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.

What formulation is right for me?

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.

7048-AD

Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl and Glycerol.
Shipping The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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.

Assay Procedure

Materials
  • Assay Buffer: 50 mM HEPES, pH 7.5
  • Recombinant Human Adenosine Deaminase/ADA (rhADA) (Catalog # 7048-AD)
  • Substrate: Adenosine (Sigma, Catalog # A9251), 10 mM stock in deionized water
  • UV plate (Costar, Catalog # 3635)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Thaw the Substrate stock and then warm it at 37 °C for 20 minutes. Mix well after incubation.
  2. Dilute rhADA to 0.1 ng/μL in Assay Buffer.
  3. Dilute Substrate to 200 μM in Assay Buffer.
  4. Load into a plate 50 μL of 0.1 ng/μL rhADA and start the reaction by adding 50 μL of 200 μM Substrate. For Substrate Blanks, load 50 μL of Assay Buffer and 50 μL of 200 μM Substrate.
  5. Read plate at a wavelength of 265 nm (bottom read) in kinetic mode for 5 minutes.
  6. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

Adjusted Vmax* (OD/min) x -1 x well volume (L) x 1012 pmol/mol
ext. coeff** (M-1cm-1) x path corr.*** (cm) x amount of enzyme (µg)

     *Adjusted for Substrate Blank 
     **Using the extinction coefficient 8500 M-1cm-1 
     ***Using the path correction 0.32 cm
     Note: the output of many spectrophotometers is in mOD Per Well:
  • rhADA: 0.005 μg
  • Substrate: 100 μM
Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Adenosine Deaminase/ADA

Adenosine Deaminase (ADA, adenosine aminohydrolase) is one of the key enzymes of purine nucleotide catabolism. It catalyses the hydrolytic deamination of adenosine and deoxy‑adenosine to inosine and deoxyinosine (1, 2). ADA is expressed in virtually all tissues and is expressed at high levels in T-lymphocytes. Adenosine Deaminase deficiency can cause a form of SCID (severe combined immunodeficiency) and lymphopenia in both B- and T-cell lineages (3, 4). ADA can be used as a sensitive diagnostic marker for tuberculous pleuritis (5). Although it is primarily a cytosolic enzyme, ADA is known to be a positive regulator of T-cell co‑activation due to its binding to CD26 at the cell surface. The interaction of ADA with CD26 regulates lymphocyte-epithelial cell adhesion (6).

References
  1. Wolfenden, R.V. et al. (1969) Biochemistry 6:2412.
  2. Lowenstein, J.M. (1972) Physiol. Rev. 52:382.
  3. Giblett, E.R. et al. (1972) Lancet 2:1067.
  4. Coleman, M.S. et al. (1978) J. Biol. Chem. 253:1619.
  5. Baba, K. et al. (2008) PLoS ONE 3:e2788.
  6. Gines, S. et al. (2002) Biochem. J. 361:203.
Long Name
Adenosine Aminohydrolase
Entrez Gene IDs
100 (Human); 11486 (Mouse); 24165 (Rat)
Alternate Names
ADA; ADA1; adenine deaminase; Adenosine aminohydrolase; Adenosine Deaminase; EC 3.5.4.4

Citation for Recombinant Human Adenosine Deaminase/ADA 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

  1. Autocrine Adenosine regulates tumor polyfunctional CD73+CD4+ effector T cells devoid of immune checkpoints
    Authors: N Gourdin, M Bossennec, C Rodriguez, S Vigano, C Machon, C Jandus, D Bauché, J Faget, I Durand, N Chopin, O Tredan, JC Marie, B Dubois, J Guitton, P Romero, C Caux, C Ménétrier-
    Cancer Res., 2018;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay

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