Recombinant Human Carbonic Anhydrase I Protein, CF

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Product Details

Summary
Reactivity HuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

Order Details

Recombinant Human Carbonic Anhydrase I Protein, CF Summary

Details of Functionality
Measured by its esterase activity. The specific activity is >10 pmol/min/µg, as measured under the described conditions.
Source
E. coli-derived human Carbonic Anhydrase I/CA1 protein
Ala2-Phe261, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Ala2
Protein/Peptide Type
Recombinant Enzymes
Gene
CA1
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

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
30 kDa, reducing conditions
Publications
Read Publication using
2180-CA 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, -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 under reducing conditions and visualized by silver stain
Assay Procedure
  • Assay Buffer: 12.5 mM Tris, 75 mM NaCl, pH 7.5
  • Recombinant Human Carbonic Anhydrase I/CA1 (rhCA1) (Catalog # 2180-CA)
  • Substrate: 4-Nitrophenyl acetate (4-NPA) (Sigma, Catalog # N8130), 100 mM stock in acetone
  • 96-well Clear Plate (Costar, Catalog # 92592)
  • Plate reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rhCA1 to 100 ng/µL in Assay Buffer.
  2. Dilute Substrate to 2 mM in Assay Buffer.
  3. Load 50 µL of 100 ng/µL rhCA1 into a plate, and start the reaction by adding 50 µL of 2 mM Substrate.
  4. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of Substrate.
  5. Read at a wavelength of 400 nm (bottom read) in kinetic mode for 5 minutes.
  6. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

Adjusted Vmax* (OD/min) x Conversion Factor** (pmol/OD)
amount of enzyme (µg)

     *Adjusted for Substrate Blank
     **Derived using calibration standard 4-Nitrophenol (Sigma, Catalog # 241326).

Per Well:
  • rhCA1: 5 µg
  • Substrate: 1 mM

Notes

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

Alternate Names for Recombinant Human Carbonic Anhydrase I Protein, CF

  • CA1
  • CA-I
  • Car1
  • Carbonate dehydratase I
  • carbonic anhydrase 1
  • Carbonic anhydrase B
  • Carbonic Anhydrase I
  • carbonic anhydrase ICAB
  • carbonic dehydratase
  • EC 4.2.1.1

Background

Carbonic Anhydrase (CA) catalyzes the reversible reaction of CO2 + H2O = HCO3- + H+, which is fundamental to many processes such as respiration, renal tubular acidification and bone resorption (1). Topics in a CA meeting (6th International Conference on the CAs, June 20-25, 2003, Slovakia) ranged from use of CAs as markers for tumor and hypoxia in clinic, as nutritional supplement in milk, and as a tool for CO2 removal and mosquito control in industry. CA1 is a cytosolic enzyme with the highest levels in erythrocytes and is a very early marker for erythroid differentiation (2). The activity of CA1 can also be measured by its ability to catalyze the reaction CO2 + H2O → HCO3- + H+, using a published method (3).

  1. Hewett-Emmett, D. and R.E. Tashian (1996) Mol. Phylogenet. Evol. 5:50.
  2. Sly, W.S. and P.Y. Hu (1995) Annu. Rev. Biochem. 64:375.
  3. Wilbur, K.M. and N.G. Anderson (1948) J. Biol. Chem. 176:147.

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Publications for Carbonic Anhydrase I/CA1 (2180-CA)(1)

We have publications tested in 1 confirmed species: Human.

We have publications tested in 1 application: Bioassay.


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Bioinformatics

Gene Symbol CA1
Uniprot