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Human Carbonic Anhydrase XIV / CA14 ELISA Pair Set

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Materials provided
Capture Ab:0.5 mg/mL of rabbit anti-CA14 monoclonal antibody, Dilute to a working concentration of 2 μg/mL in CBS before coating.
Detection Ab:0.5 mg/mL rabbit anti-CA14 polyclonal antibody conjugated to horseradish-peroxidase (HRP). Dilute to working concentration of 0.25 μg/mL in detection antibody dilution buffer before use.
Standard:Each vial contains 235 ng of recombinant CA14. Reconstitute with 1 mL detection antibody dilution buffer. After reconstitution, store at -20℃ to -70℃ in a manual defrost freezer. A seven-point standard curve usi ng 2-fold serial dilutions in sample dilution buffer, and a high standard of 8 ng/mL is recommended.
Sensitivity
The minimum detectable dose of Human Carbonic Anhydrase XIV / CA14 was determined to be approximately 125 pg/ml. This is defined as at least three times standard deviations above the mean optical density of 10 replicates of the zero standard.
Principle of the product
The Human Carbonic Anhydrase XIV / CA14 ELISA Pair Set is for the quantitative determination of Human Carbonic Anhydrase XIV / CA14.
This ELISA Pair Set contains the basic components required for the development of sandwich ELISAs.
The Sino Biological ELISA Pair Set is a solid phase sandwich ELISA (Enzyme-Linked Immunosorbent Assay). It utilizes a monoclonal antibody specific for Carbonic Anhydrase XIV / CA14 coated on a 96-well plate. Standards and samples are added to the wells, and any Carbonic Anhydrase XIV / CA14 present binds to the immobilized antibody. The wells are washed and a horseradish peroxidase conjugated rabbit anti-Carbonic Anhydrase XIV / CA14 polyclonal antibody is then added, producing an antibody-antigen-antibody "sandwich". The wells are again washed and TMB substrate solution is loaded, which produces color in proportion to the amount of Carbonic Anhydrase XIV / CA14 present in the sample. To end the enzyme reaction, the stop solution is added and absorbances of the microwell are read at 450 nm.
Storage
Capture Antibody: Aliquot and store at -20℃ to -70℃ for up to 6 months from date of receipt. Avoid repeated freeze-thaw cycles.
Detection Antibody: Protect it from prolonged exposure to light. Aliquot and store at -20℃ to -70℃ and for up to 6 months from date of receipt. Avoid repeated freeze-thaw cycles.
Standard: Store lyophilized Standard at -20℃ to -70℃ for up to 6 months from date of receipt. Aliquot and store the reconstituted Standard at -20℃ to -70℃ for up to 1 month. Avoid repeated freeze-thaw cycles.
Image
Human Carbonic Anhydrase XIV / CA14 ELISA standard curve

Background

The carbonic anhydrases (or carbonate dehydratases) are classified as metalloenzyme for its zinc ion prosthetic group and form a family of enzymes that catalyze the rapid interconversion of carbon dioxide and water to bicarbonate and protons, a reversible reaction that takes part in maintaining acid-base balance in blood and other tissues. The carbonic anhydrasekl (CA) family consists of at least 11 enzymatically active members and a few inactive homologous proteins. CAXIV is a member of CA family that showed an overall similarity of 29–46% to other active CA isozymes. The highest percentage similarity was with a transmembrane CA isoform, CAXII. The CAXIV was found high concentrations in human heart, brain, liver, and skeletal muscle but lower in the colon, small intestine, urinary bladder, and kidney. No CAXIV mRNA was seen in the salivary gland and pancreas. CAXIV is a likely candidate for the extracellular CA postulated to have an important role in modulating excitatory synaptic transmission in brain.

References
  • Lehtonen J, et al. (2004) Characterization of CA XIII, a Novel Member of the Carbonic Anhydrase Isozyme Family. The Journal of Biological Chemistry. 279: 2719-27.
  • Lindskog S. (1997) Structure and mechanism of carbonic anhydrase. Pharmacology & Therapeutics. 74(1):1-20.
  • Parkkila S, et al. (2001) Expression of membrane-associated carbonic anhydrase XIV on neurons and axons in mouse and human brain. PNAS. 98(4): 1918-23.
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