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Reports of Biochemistry and Molecular Biology
rbmb.net
Basic Sciences
http://rbmb.net
1
admin
2322-3480
2322-3480
10.61882/rbmb
en
jalali
1400
5
1
gregorian
2021
8
1
10
2
online
1
fulltext
en
Accurate and Precise Protocol to Estimate the Activity of Peroxiredoxin Enzyme
بیوشیمی
Biochemistry
مقالات اصلی
Original Article
<div style="text-align: justify;"><strong><em>Background:</em></strong> Accurate estimation of Peroxiredoxin Enzyme (Prx) activity poses many complications and interferences. The present protocol is free of interference and provides an effective alternative for the assessment of peroxide with high sensitivity. The assay can be used in clinical pathology laboratories since it is simple, rapid, and inexpensive. The systematic reagent consisted of AFS/ASA which acted as a sensitive probe for peroxide.<br>
<br>
<strong><em>Methods:</em></strong> Prx activity was estimated by incubating samples in suitable concentrations of 1,4-dithio-DL-threitol (DTT) and hydrogen peroxide (H2O2) or t-Butyl hydroperoxide (t-BOOH), as the substrates. The enzymatic reaction was inhibited after incubation with a working reagent containing ammonium ferrous sulfate (AFS) and aminosalicylic acid (ASA).<br>
<br>
<strong><em>Results:</em></strong> Residual peroxide reacted with the working solution to form a brown-colored ferriaminosalicylate (FAS) complex with a maximum absorbance (λmax) of 425 nm. This protocol used sodium azide (NaN3) to eliminate catalase interference and avoided using high concentrations of a strong acid to inhibit the Prx reaction.<br>
<br>
<strong><em>Conclusions:</em></strong> We concluded that the new protocol produced the same efficacy as the reference method since a strong correlation coefficient of comparison (r> 0.99) was found between both the FAS and ferrithiocyanate method.</div>
Amino Salicylic Acid, Ammonium Ferrous Sulfate, Dithiothreitol, Peroxiredoxin, T-Butyl Hydroperoxide.
156
163
http://rbmb.net/browse.php?a_code=A-10-603-1&slc_lang=en&sid=1
Thulfeqar
Hamza
100319475328460012591
100319475328460012591
No
Chemistry Dept., College of Science, University of Babylon, Iraq & Pathological Analysis Department, Al-Mustaqbal University College, Hilla City, Babylon Governorate, Iraq.
Mahmoud Hussein
Hadwan
sci.mahmood.hadwan@uobabylon.edu.iq.
100319475328460012592
100319475328460012592
Yes
Pathological Analysis Department, Al-Mustaqbal University College, Hilla City, Babylon Governorate, Iraq.