Carbonic anhydrase III in equine tissues and sera determined by a highly sensitive enzyme-immunoassay.
Abstract: A sensitive sandwich enzyme immunoassay (EIA) for measuring equine carbonic anhydrase III (CA-III) was established using a microplate as a solid-phase and peroxidase as a labelling enzyme. The assay can detect concentrations as low as 5 ng/ml using 20 microliters of sample sera. Within-run coefficients of variation obtained using standard equine CA-III were less than 5 per cent. CA-III levels in equine serum ranged from 5 to 50 ng/ml (n = 370), and apparently abnormal levels of CA-III from 100 to 1900 ng/ml (n = 27) were observed. The concentrations of immunoreactive CA-III in the extracts of various equine tissues were also determined; it was present at high concentrations in skeletal muscle and liver and to a much lesser extent in the thymus. Other tissues contained much smaller amounts.
Publication Date: 1990-07-01 PubMed ID: 2120033DOI: 10.1111/j.2042-3306.1990.tb04262.xGoogle Scholar: Lookup
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Summary
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This research paper develops and tests an enzyme immunoassay (EIA) for detecting equine carbonic anhydrase III (CA-III), an enzyme found in horses, and measures its concentration in various equine tissues. The novel EIA proves to be sensitive and capable of detecting CA-III concentrations as low as 5 ng/ml, making it an efficient tool for assessing carbonic anhydrase III levels in horses.
Development of the Enzyme Immunoassay (EIA)
- The research team established a sandwich enzyme immunoassay (EIA), which is a highly sensitive and specific method used to detect and quantify substances such as enzymes, hormones, or antibodies.
- In the study, this assay was used to measure levels of equine carbonic anhydrase III (CA-III), an enzyme found in horses’ tissues and serum.
- The EIA used a microplate as a solid-phase and peroxidase as a labelling enzyme, which is the enzyme used to mark the target enzyme (in this case, carbonic anhydrase III) for detection.
- Within-run coefficients of variation obtained using standard equine CA-III were less than 5 per cent, indicating the high precision and reproducibility of the EIA test.
Detection and Measurement of CA-III
- The new EIA can detect CA-III concentrations as low as 5 ng/ml with only 20 microliters of sample sera, making it a cost-effective and efficient method of enzyme detection for equine veterinary diagnostics.
- The levels of CA-III in equine serum were found to range from 5 to 50 ng/ml for the majority (n = 370) of samples and were significantly higher in a smaller sample group (n = 27), in which CA-III levels ranged from 100 to 1900 ng/ml. These differences highlight the potential variations in CA-III levels amongst horse populations.
Assessment of CA-III in Equine Tissues
- Apart from serum samples, the concentrations of immunoreactive CA-III in the extracts of various equine tissues were also determined.
- The enzyme was found at high concentrations in skeletal muscle and liver, indicating its key role in these tissues’ functions.
- To a much lesser extent, CA-III presence was also detected in the thymus, while it was found in much smaller amounts in other tissues.
- This tissue-specific distribution of CA-III may give insight into the functional role of this enzyme in horses.
Cite This Article
APA
Nishita T, Matsushita H.
(1990).
Carbonic anhydrase III in equine tissues and sera determined by a highly sensitive enzyme-immunoassay.
Equine Vet J, 22(4), 247-250.
https://doi.org/10.1111/j.2042-3306.1990.tb04262.x Publication
Researcher Affiliations
- Laboratory of Veterinary Physiology, School of Veterinary Medicine, Azabu University, Kanagawa, Japan.
MeSH Terms
- Animals
- Carbonic Anhydrases / analysis
- Carbonic Anhydrases / blood
- Erythrocytes / enzymology
- Female
- Horses / blood
- Horses / metabolism
- Immunoenzyme Techniques
- Liver / enzymology
- Male
- Muscles / enzymology
- Predictive Value of Tests
- Thymus Gland / enzymology
Citations
This article has been cited 3 times.- Nishita T, Tomita Y, Yorifuji D, Orito K, Ochiai H, Arishima K. Purification of chicken carbonic anhydrase isozyme-III (CA-III) and its measurement in White Leghorn chickens. Acta Vet Scand 2011 Nov 26;53(1):63.
- Nishita T, Oshige H, Matsushita H, Kano Y, Asari M. The immunohistolocalization of carbonic anhydrase III in the submandibular gland of rats and hamsters. Histochem J 1989 Jan;21(1):8-14.
- Asari M, Igarashi S, Sasaki K, Amasaki T, Nishita T, Amasaki H. Immunocytochemical localisation of the carbonic anhydrase III in the rat parotid gland. J Anat 1991 Dec;179:9-14.
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