Zymoblot
Zymoblot is the fastest available
Background
Physiological phenomena whether at the cellular or molecular level in
Procedure
As little as 1µl, or less, of a sample is enough to detect enzyme activity by the zymoblot technique as the coloured product being
The technique has advantages not shared by any other technique. Samples to be analysed by zymoblot require no dialysis (a process that may take days) as washing blots in Tris-buffered saline (TBS) before marinating them in the reaction mixture does remove inhibitors. In contrast to the other techniques where samples are assayed individually, samples to be analysed by zymoblot are spotted on the same blot and enzyme activity is assayed with the same reaction mixture at the same time minimising experimental errors and allowing quick qualitative comparisons. Additionally, several enzymes can be assayed in a sequential order on the same blot. That is to say, if a blot proves negative for a certain enzyme, it can be washed in TBS and reused for another enzyme and so on until a positive reaction for an enzyme is obtained. Unlike wet assays (e.g. colorimetry), results obtained by the zymoblot are always in a recorded from. This allows zymoblots to be carried out in one place, where a densitometer may not be available, and taken or sent to another place to be quantitatively assayed by densitometry. Taking a zymoblot in a researcher's wallet to a meeting facilitates discussions and exchange of ideas with other scientists. While immunologically-based enzyme assays[3] which uses enzyme specific antibodies to directly detect enzymes suffers from the major disadvantage of measuring the "total enzyme content" and not the total enzyme activity, zymoblot, being not a serological technique, uses no antibodies and measures enzyme activity as it detects only the active (functional) portion of the total enzyme content in a sample.
Zymoblot is an end-point type of assay where an enzyme reaction is allowed to proceed for a fixed period of time before being stopped by rapid elimination of its specific
Applications
The Zymoblot technique is simpler, cheaper, more reliable and less time-consuming than all known procedures for enzyme assays. It is probably the quickest available technique to detect enzyme activity in any biological or even non-biological specimen. The technique is highly competitive in price with all commercially available kits. Such advantages should qualify the Zymoblot technique for wide potential uses in
Qualitative zymoblot is of great potential use in diagnosis of human, animal and plant
References
- ^ doi:10.1139/m93-077.Wagih, E. E. and Fletcher, J. (1993). Zymoblot, a new microtechnique used to detect enzyme activity in spiroplasmas and bacteria. Canadian Journal of Microbiology 39: 543- 547.
- ^ Dixon, M. and Webb, E.C. (1979). Enzymes. 3rd ed. Longman Group Ltd. 1116pp.
- ^ a b Wiseman, A. [ed.] (1983). Topics in enzyme and fermentation Biotechnology 7. Ellis Horwood Ltd. 314pp.
- ^ Wagih, E.E. and Wagih, M.E. (1996). The Zymoblot Technique: Potential in Plant Physiology. Proc. 2nd Asia-Pacific Conference on Plant Physiology, Shah Alam, Kuala Lumpur, Malaysia, 20–22 August 1996.
- ^ Wagih, M. E., Onwueme, I. C. and Wagih, E. E. (1996). Detection of differential peroxidase gene expression in taro (Colocasia esculenta L. Scotts) using the zymoblot technique. In the 2nd International Crop Science Congress. New Delhi, India, 17–24 November 1996.
- ^ Wagih, E. E. and Wagih, M. E. (1997). Quantitative Zymoblot and Proteinblot Techniques and their Use in Monitoring Total Enzyme and Soluble Protein Alterations in Plant Virus Infections Proc. 1st All Africa Crop Science Conference, Pretoria, The Republic of South Africa, 13–17 January 1997.
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