371:. When an enzyme is mixed with a large excess of the substrate, the enzyme-substrate intermediate builds up in a fast initial transient. Then the reaction achieves a steady-state kinetics in which enzyme substrate intermediates remains approximately constant over time and the reaction rate changes relatively slowly. Rates are measured for a short period after the attainment of the quasi-steady state, typically by monitoring the accumulation of product with time. Because the measurements are carried out for a very short period and because of the large excess of substrate, the approximation that the amount of free substrate is approximately equal to the amount of the initial substrate can be made. The initial rate experiment is the simplest to perform and analyze, being relatively free from complications such as back-reaction and enzyme degradation. It is therefore by far the most commonly used type of experiment in enzyme kinetics.
621:(MST) measures the size, charge and hydration entropy of molecules/substrates at equilibrium. The thermophoretic movement of a fluorescently labeled substrate changes significantly as it is modified by an enzyme. This enzymatic activity can be measured with high time resolution in real time. The material consumption of the all optical MST method is very low, only 5 μl sample volume and 10nM enzyme concentration are needed to measure the enzymatic rate constants for activity and inhibition. MST allows analysts to measure the modification of two different substrates at once (
31:
690:
491:
422:
377:. In these experiments, the kinetic parameters are determined from expressions for the species concentrations as a function of time. The concentration of the substrate or product is recorded in time after the initial fast transient and for a sufficiently long period to allow the reaction to approach equilibrium. Progress curve experiments were widely used in the early period of enzyme kinetics, but are less common now.
1430:
560:
677:. Although this approach can need a lot of material, its sensitivity can be increased by labelling the substrates/products with a radioactive or fluorescent tag. Assay sensitivity has also been increased by switching protocols to improved chromatographic instruments (e.g. ultra-high pressure liquid chromatography) that operate at pump pressure a few-fold higher than HPLC instruments (see
570:
is the measurement of the heat released or absorbed by chemical reactions. These assays are very general, since many reactions involve some change in heat and with use of a microcalorimeter, not much enzyme or substrate is required. These assays can be used to measure reactions that are impossible to
533:
after absorbing light of a different wavelength. Fluorometric assays use a difference in the fluorescence of substrate from product to measure the enzyme reaction. These assays are in general much more sensitive than spectrophotometric assays, but can suffer from interference caused by impurities and
405:
or pH jump, and the return to equilibrium is monitored. The analysis of these experiments requires consideration of the fully reversible reaction. Moreover, relaxation experiments are relatively insensitive to mechanistic details and are thus not typically used for mechanism identification, although
350:
is 100% × (specific activity of enzyme sample / specific activity of pure enzyme). The impure sample has lower specific activity because some of the mass is not actually enzyme. If the specific activity of 100% pure enzyme is known, then an impure sample will have a lower specific activity, allowing
753:
found in the hot springs are stable up to 100 °C. However, the idea of an "optimum" rate of an enzyme reaction is misleading, as the rate observed at any temperature is the product of two rates, the reaction rate and the denaturation rate. If you were to use an assay measuring activity for one
582:
is the emission of light by a chemical reaction. Some enzyme reactions produce light and this can be measured to detect product formation. These types of assay can be extremely sensitive, since the light produced can be captured by photographic film over days or weeks, but can be hard to quantify,
363:
measure either the consumption of substrate or production of product over time. A large number of different methods of measuring the concentrations of substrates and products exist and many enzymes can be assayed in several different ways. Biochemists usually study enzyme-catalysed reactions using
786:
concentration increases the rate of reaction (enzyme activity). However, enzyme saturation limits reaction rates. An enzyme is saturated when the active sites of all the molecules are occupied most of the time. At the saturation point, the reaction will not speed up, no matter how much additional
732:
All enzymes work within a range of temperature specific to the organism. Increases in temperature generally lead to increases in reaction rates. There is a limit to the increase because higher temperatures lead to a sharp decrease in reaction rates. This is due to the denaturating (alteration) of
649:
most frequently used in these assays are C, P, S and I. Since radioactive isotopes can allow the specific labelling of a single atom of a substrate, these assays are both extremely sensitive and specific. They are frequently used in biochemistry and are often the only way of measuring a specific
326:
An active site titration process can be done for the elimination of errors arising from differences in cultivation batches and/or misfolded enzyme and similar issues. This is a measure of the amount of active enzyme, calculated by e.g. titrating the amount of active sites present by employing an
609:
measures the product of weight-averaged molar mass and concentration of macromolecules in solution. Given a fixed total concentration of one or more species over the measurement time, the scattering signal is a direct measure of the weight-averaged molar mass of the solution, which will vary as
537:
An example of these assays is again the use of the nucleotide coenzymes NADH and NADPH. Here, the reduced forms are fluorescent and the oxidised forms non-fluorescent. Oxidation reactions can therefore be followed by a decrease in fluorescence and reduction reactions by an increase. Synthetic
745:
that stabilize the three-dimensional structure of the enzyme active site. The "optimum" temperature for human enzymes is usually between 35 and 40 °C. The average temperature for humans is 37 °C. Human enzymes start to denature quickly at temperatures above 40 °C. Enzymes from
383:. In these experiments, reaction behaviour is tracked during the initial fast transient as the intermediate reaches the steady-state kinetics period. These experiments are more difficult to perform than either of the above two classes because they require specialist techniques (such as
311:
in a given amount of time (minutes) under given conditions per milligram of total proteins. Specific activity is equal to the rate of reaction multiplied by the volume of reaction divided by the mass of total protein. The SI unit is katal/kg, but a more practical unit is μmol/(mg*min).
306:
The specific activity of an enzyme is another common unit. This is the activity of an enzyme per milligram of total protein (expressed in μmol min mg). Specific activity gives a measurement of enzyme purity in the mixture. It is the micro moles of product formed by an
450:
assays, you follow the course of the reaction by measuring a change in how much light the assay solution absorbs. If this light is in the visible region you can actually see a change in the color of the assay, and these are called
331:, or μmol product per second per μmol of active enzyme, can be calculated from the specific activity. The turnover number can be visualized as the number of times each enzyme molecule carries out its catalytic cycle per second.
146:
610:
complexes form or dissociate. Hence the measurement quantifies the stoichiometry of the complexes as well as kinetics. Light scattering assays of protein kinetics is a very general technique that does not require an enzyme.
297:
An increased amount of substrate will increase the rate of reaction with enzymes, however once past a certain point, the rate of reaction will level out because the amount of active sites available has stayed constant.
282:
Enzyme activity as given in katal generally refers to that of the assumed natural target substrate of the enzyme. Enzyme activity can also be given as that of certain standardized substrates, such as
754:
second, it would give high activity at high temperatures, however if you were to use an assay measuring product formation over an hour, it would give you low activity at these temperatures.
205:
633:
Discontinuous assays are when samples are taken from an enzyme reaction at intervals and the amount of product production or substrate consumption is measured in these samples.
766:
and have specific ranges of activity. All have an optimum pH. The pH can stop enzyme activity by denaturating (altering) the three-dimensional shape of the enzyme by breaking
506:
Even when the enzyme reaction does not result in a change in the absorbance of light, it can still be possible to use a spectrophotometric assay for the enzyme by using a
253:
229:
174:
969:
438:
Continuous assays are most convenient, with one assay giving the rate of reaction with no further work necessary. There are many different types of continuous assays.
510:. Here, the product of one reaction is used as the substrate of another, easily detectable reaction. For example, figure 1 shows the coupled assay for the enzyme
294:(MCU). The units GDU and MCU are based on how fast one gram of the enzyme will digest gelatin or milk proteins, respectively. 1 GDU approximately equals 1.5 MCU.
482:
using NADH as a substrate could therefore be assayed by following the decrease in UV absorbance at a wavelength of 340 nm as it consumes the coenzyme.
697:
Several factors effect the assay outcome and a recent review summarizes the various parameters that needs to be monitored to keep an assay up and running.
98:
2018:
327:
irreversible inhibitor. The specific activity should then be expressed as μmol min mg active enzyme. If the molecular weight of the enzyme is known, the
538:
substrates that release a fluorescent dye in an enzyme-catalyzed reaction are also available, such as 4-methylumbelliferyl-β-D-galactoside for assaying
1233:
678:
2071:
1305:
Daniel RM, Peterson ME, Danson MJ, et al. (January 2010). "The molecular basis of the effect of temperature on enzyme activity".
910:
1910:
2158:
774:. Most enzymes function between a pH of 6 and 8; however pepsin in the stomach works best at a pH of 2 and trypsin at a pH of 8.
934:"The mechanism distinguishability problem in biochemical kinetics: The single-enzyme, single-substrate reaction as a case study"
2153:
670:
933:
85:
Enzyme activity is a measure of the quantity of active enzyme present and is thus dependent on various physical conditions,
1486:
1387:"The influence of macromolecular crowding and macromolecular confinement on biochemical reactions in physiological media"
1081:
Todd MJ, Gomez J (September 2001). "Enzyme kinetics determined using calorimetry: a general assay for enzyme activity?".
1797:
625:) if both substrates are labeled with different fluorophores. Thus substrate competition experiments can be performed.
999:
515:
499:
467:
463:
2008:
1988:
1765:
880:
825:
447:
1969:
17:
397:. In these experiments, an equilibrium mixture of enzyme, substrate and product is perturbed, for instance by a
1434:
1284:
418:, where samples are taken, the reaction stopped and then the concentration of substrates/products determined.
1998:
1903:
181:
2028:
2046:
914:
713:. Typical enzymes are active in salt concentrations of 1-500 mM. As usual there are exceptions such as the
665:
Chromatographic assays measure product formation by separating the reaction mixture into its components by
873:
2194:
2189:
2145:
2051:
1979:
267:
s (mole per second), but this is an excessively large unit. A more practical and commonly used value is
2003:
1993:
1983:
1945:
830:
674:
618:
514:, which can be assayed by coupling its production of glucose-6-phosphate to NADPH production, using
2179:
2023:
1896:
850:
783:
1350:
Cowan DA (1997). "Thermophilic proteins: stability and function in aqueous and organic solvents".
705:
Most enzymes cannot tolerate extremely high salt concentrations. The ions interfere with the weak
236:
212:
157:
1872:
1479:
808:
606:
2094:
2056:
1441:
343:
is the concentration of substrate disappearing (or product produced) per unit time (mol L s).
2013:
1660:
1628:
654:
1032:"A fast, miniaturised in-vitro assay developed for quantification of lipase enzyme activity"
459:, a redox assay using a tetrazolium dye as substrate is an example of a colorimetric assay.
2061:
1940:
1830:
1825:
1782:
1732:
1623:
1602:
1451:
1186:
1137:
918:
804:
650:
reaction in crude extracts (the complex mixtures of enzymes produced when you lyse cells).
646:
8:
1974:
1965:
1787:
1755:
1597:
1565:
1190:
1141:
598:, this is found in fireflies and naturally produces light from its substrate luciferin.
2199:
1960:
1815:
1807:
1792:
1760:
1740:
1648:
1472:
1209:
1174:
1106:
1058:
1031:
961:
894:
875:
845:
1363:
539:
2115:
1852:
1408:
1367:
1332:
1276:
1214:
1155:
1098:
1063:
995:
953:
579:
1110:
965:
141:{\displaystyle \mathrm {a} =\mathrm {n} _{\text{t}}=\mathrm {r} \times \mathrm {V} }
2184:
2130:
1772:
1638:
1524:
1398:
1359:
1322:
1314:
1268:
1204:
1194:
1145:
1090:
1053:
1043:
945:
889:
874:
Nomenclature
Committee of the International Union of Biochemistry (NC-IUB) (1979).
742:
591:
590:
by enzymatic chemiluminescence (ECL) is a common method of detecting antibodies in
398:
384:
320:
54:
1272:
1048:
2099:
1923:
1888:
1840:
1820:
931:
855:
328:
50:
2038:
1955:
1862:
1857:
1777:
949:
666:
544:
479:
410:
Enzyme assays can be split into two groups according to their sampling method:
319:(the capability of enzyme to be processed), at a specific (usually saturating)
1126:"Protein-binding assays in biological liquids using microscale thermophoresis"
2173:
2135:
2125:
2066:
1745:
1558:
800:
796:
771:
642:
402:
1199:
2120:
1950:
1677:
1653:
1643:
1548:
1536:
1529:
1412:
1403:
1386:
1336:
1280:
1218:
1159:
1102:
1094:
1067:
957:
747:
622:
526:
388:
1371:
1254:
30:
1835:
1701:
1582:
1577:
1553:
1446:
706:
689:
567:
268:
264:
74:
70:
1932:
1867:
1710:
1706:
1682:
1611:
1592:
1318:
1150:
1125:
767:
738:
714:
595:
587:
530:
511:
495:
42:
1327:
1722:
1665:
1587:
1514:
1495:
820:
534:
the instability of many fluorescent compounds when exposed to light.
456:
1877:
1672:
1633:
1618:
1541:
1519:
1256:
721:
710:
583:
because not all the light released by a reaction will be detected.
1257:"Improving Sensitivity in Liquid Chromatography-Mass Spectrometry"
693:
A pressure chamber for measuring enzyme activity at high pressure.
1919:
1750:
1715:
1695:
1572:
1509:
1255:
Churchwell, M; Twaddle, N; Meeker, L; Doerge, D. (October 2005).
787:
substrate is added. The graph of the reaction rate will plateau.
750:
734:
426:
421:
283:
932:
Schnell, S.; Chappell, M.J.; Evans, N.D.; Roussel, M.R. (2006).
490:
73:
amounts, as with any other chemical, or in terms of activity in
1429:
548:
308:
66:
46:
653:
Radioactivity is usually measured in these procedures using a
559:
414:, where the assay gives a continuous reading of activity, and
2081:
1236:[Optically generated thermophoresis for bioanalysis]
717:
475:
471:
360:
276:
260:
1464:
1014:
994:. Vol. 4. New York: Academic Press. pp. 2066–72.
323:
concentration, and is usually constant for a pure enzyme.
272:
351:
purity to be calculated and then getting a clear result.
763:
275:
min (micromole per minute). 1 U corresponds to 16.67
239:
215:
184:
160:
101:
1304:
1234:"Optisch erzeugte Thermophorese für die Bioanalytik"
1036:
Journal of Enzyme
Inhibition and Medicinal Chemistry
679:
High-performance liquid chromatography#Pump pressure
645:
into substrates or its release from substrates. The
737:structure resulting from the breakdown of the weak
462:UV light is often used, since the common coenzymes
1918:
673:(HPLC), but can also use the simpler technique of
247:
223:
199:
168:
140:
27:Laboratory method for measuring enzymatic activity
1231:
1175:"Why molecules move along a temperature gradient"
2171:
911:"How Many? A Dictionary of Units of Measurement"
641:Radiometric assays measure the incorporation of
989:
807:of enzyme reactions, through an effect called
542:or 4-methylumbelliferyl-butyrate for assaying
92:It is calculated using the following formula:
1904:
1480:
387:of caged compounds) or rapid mixing (such as
613:
207:= Moles of substrate converted per unit time
1019:. Totowa NJ: Humana Press. pp. 85–110.
684:
1911:
1897:
1487:
1473:
1123:
902:
290:(GDU), or milk proteins, then measured in
49:activity. They are vital for the study of
1402:
1384:
1326:
1208:
1198:
1172:
1149:
1080:
1057:
1047:
893:
727:
406:they can be under appropriate conditions.
1349:
814:
688:
558:
489:
420:
29:
1225:
908:
628:
200:{\displaystyle \mathrm {n} _{\text{t}}}
14:
2172:
2154:Photoactivated localization microscopy
2072:Protein–protein interaction prediction
1378:
1117:
1029:
1015:Passonneau, J.V.; Lowry, O.H. (1993).
671:high-performance liquid chromatography
529:is when a molecule emits light of one
334:
34:Beckman DU640 UV/Vis spectrophotometer
1892:
1468:
1017:Enzymatic Analysis. A Practical Guide
700:
441:
1232:Baaske P, Wienken C, Duhr S (2009).
1166:
790:
777:
433:
391:, quenched flow or continuous flow).
301:
65:The quantity or concentration of an
2029:Freeze-fracture electron microscopy
601:
574:
24:
1798:Fluorescence in situ hybridization
909:Rowlett, Russ (23 November 1998).
895:10.1111/j.1432-1033.1979.tb13116.x
660:
354:
315:Specific activity is a measure of
241:
217:
187:
162:
134:
126:
112:
103:
80:
25:
2211:
1422:
1173:Duhr S, Braun D (December 2006).
516:glucose-6-phosphate dehydrogenase
500:glucose-6-phosphate dehydrogenase
2009:Isothermal titration calorimetry
1989:Dual-polarization interferometry
1766:Oral and maxillofacial pathology
1442:"Assays Protocols - OpenWetWare"
1428:
1124:Wienken CJ; et al. (2010).
881:European Journal of Biochemistry
826:Fluorescein diacetate hydrolysis
757:
594:. Another example is the enzyme
1343:
1298:
1287:from the original on 2022-01-20
1248:
972:from the original on 2024-02-26
554:
521:
60:
1074:
1030:Menden, Ariane (26 Jul 2019).
1023:
1008:
983:
925:
867:
762:Most enzymes are sensitive to
636:
429:holder in a spectrophotometer.
381:Transient kinetics experiments
13:
1:
1999:Chromatin immunoprecipitation
1494:
1364:10.1016/S0300-9629(97)00004-2
1273:10.1016/j.jchromb.2005.05.037
1049:10.1080/14756366.2019.1651312
992:Methods of Enzymatic Analysis
861:
799:in a solution will alter the
2062:Protein structural alignment
2047:Protein structure prediction
1179:Proc. Natl. Acad. Sci. U.S.A
915:University of North Carolina
563:Chemiluminescence of luminol
486:Direct versus coupled assays
248:{\displaystyle \mathrm {V} }
224:{\displaystyle \mathrm {r} }
169:{\displaystyle \mathrm {a} }
7:
2146:Super-resolution microscopy
2052:Protein function prediction
1980:Peptide mass fingerprinting
1975:Protein immunoprecipitation
1261:Journal of Chromatography B
839:
474:forms, but do not in their
364:four types of experiments:
10:
2216:
950:10.1016/j.crvi.2005.09.005
876:"Units of Enzyme Activity"
669:. This is usually done by
375:Progress curve experiments
2144:
2108:
2080:
2037:
2004:Surface plasmon resonance
1994:Microscale thermophoresis
1984:Protein mass spectrometry
1946:Green fluorescent protein
1931:
1806:
1731:
1502:
1352:Comp. Biochem. Physiol. A
675:thin layer chromatography
619:Microscale thermophoresis
614:Microscale thermophoresis
470:absorb UV light in their
87:which should be specified
2024:Cryo-electron microscopy
990:Bergmeyer, H.U. (1974).
938:Comptes Rendus Biologies
851:DNase footprinting assay
685:Factors affecting assays
571:assay in any other way.
369:Initial rate experiments
2057:Protein–protein docking
1970:Protein electrophoresis
1873:Microbiological culture
1503:Principles of pathology
1200:10.1073/pnas.0603873103
809:macromolecular crowding
607:Static light scattering
425:Temperature-controlled
288:gelatin digesting units
1956:Protein immunostaining
1404:10.1074/jbc.R100005200
1095:10.1006/abio.2001.5218
728:Effects of Temperature
694:
588:horseradish peroxidase
564:
503:
430:
395:Relaxation experiments
249:
231:= Rate of the reaction
225:
201:
170:
142:
45:methods for measuring
35:
2014:X-ray crystallography
1836:Diagnostic immunology
1661:Programmed cell death
1629:Liquefactive necrosis
1130:Nature Communications
834:-Nitrophenylphosphate
815:List of enzyme assays
805:equilibrium constants
692:
655:scintillation counter
562:
493:
424:
250:
226:
202:
171:
143:
33:
1941:Protein purification
1831:Medical microbiology
1826:Transfusion medicine
1783:Immunohistochemistry
1733:Anatomical pathology
1624:Coagulative necrosis
1452:BioBricks Foundation
1437:at Wikimedia Commons
647:radioactive isotopes
629:Discontinuous assays
416:discontinuous assays
237:
213:
182:
158:
99:
69:can be expressed in
1966:Gel electrophoresis
1788:Electron microscopy
1756:Molecular pathology
1634:Gangrenous necrosis
1566:Cellular adaptation
1244:(in German): 22–24.
1191:2006PNAS..10319678D
1142:2010NatCo...1..100W
453:colorimetric assays
335:Related terminology
317:enzyme processivity
292:milk clotting units
286:, then measured in
263:, 1 katal = 1
259:The SI unit is the
2195:Clinical pathology
2190:Chemical pathology
2109:Display techniques
1961:Protein sequencing
1816:Clinical chemistry
1808:Clinical pathology
1793:Immunofluorescence
1761:Forensic pathology
1741:Surgical pathology
1649:Fibrinoid necrosis
1385:Minton AP (2001).
1319:10.1042/BJ20091254
1151:10.1038/ncomms1093
921:on 29 August 2018.
846:Restriction enzyme
701:Salt Concentration
695:
565:
504:
494:Coupled assay for
448:spectrophotometric
442:Spectrophotometric
431:
341:rate of a reaction
245:
221:
197:
166:
138:
36:
2167:
2166:
2116:Bacterial display
1886:
1885:
1853:Mass spectrometry
1433:Media related to
795:Large amounts of
791:Level of crowding
778:Enzyme Saturation
586:The detection of
580:Chemiluminescence
434:Continuous assays
412:continuous assays
302:Specific activity
255:= Reaction volume
194:
176:= Enzyme activity
119:
55:enzyme inhibition
16:(Redirected from
2207:
2131:Ribosome display
2067:Protein ontology
1913:
1906:
1899:
1890:
1889:
1773:Gross processing
1639:Caseous necrosis
1489:
1482:
1475:
1466:
1465:
1461:
1459:
1458:
1432:
1417:
1416:
1406:
1397:(14): 10577–80.
1382:
1376:
1375:
1347:
1341:
1340:
1330:
1302:
1296:
1295:
1293:
1292:
1252:
1246:
1245:
1239:
1229:
1223:
1222:
1212:
1202:
1185:(52): 19678–82.
1170:
1164:
1163:
1153:
1121:
1115:
1114:
1078:
1072:
1071:
1061:
1051:
1042:(1): 1474–1480.
1027:
1021:
1020:
1012:
1006:
1005:
987:
981:
980:
978:
977:
929:
923:
922:
917:. Archived from
906:
900:
899:
897:
871:
743:hydrogen bonding
602:Light scattering
592:western blotting
575:Chemiluminescent
385:flash photolysis
254:
252:
251:
246:
244:
230:
228:
227:
222:
220:
206:
204:
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196:
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147:
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121:
120:
117:
115:
106:
21:
2215:
2214:
2210:
2209:
2208:
2206:
2205:
2204:
2180:Protein methods
2170:
2169:
2168:
2163:
2140:
2104:
2100:Secretion assay
2076:
2033:
1927:
1917:
1887:
1882:
1841:Immunopathology
1821:Hematopathology
1802:
1727:
1498:
1493:
1456:
1454:
1440:
1425:
1420:
1383:
1379:
1348:
1344:
1303:
1299:
1290:
1288:
1253:
1249:
1237:
1230:
1226:
1171:
1167:
1122:
1118:
1079:
1075:
1028:
1024:
1013:
1009:
1002:
988:
984:
975:
973:
930:
926:
913:. Chapel Hill:
907:
903:
872:
868:
864:
856:Enzyme kinetics
842:
817:
793:
782:Increasing the
780:
760:
730:
720:and halophilic
703:
687:
663:
661:Chromatographic
639:
631:
616:
604:
577:
557:
540:β-galactosidase
524:
444:
436:
357:
355:Types of assays
337:
329:turnover number
304:
240:
238:
235:
234:
216:
214:
211:
210:
191:
186:
185:
183:
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1423:External links
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758:Effects of pH
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78:
76:
72:
68:
58:
56:
52:
48:
44:
40:
39:Enzyme assays
32:
19:
2121:mRNA display
2090:Enzyme assay
2089:
1951:Western blot
1933:Experimental
1848:Enzyme assay
1847:
1694:
1689:
1678:Karyorrhexis
1654:Myocytolysis
1644:Fat necrosis
1549:Inflammation
1537:Hemodynamics
1530:Pathogenesis
1455:. Retrieved
1445:
1394:
1390:
1380:
1355:
1351:
1345:
1310:
1306:
1300:
1289:. Retrieved
1264:
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1250:
1241:
1227:
1182:
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1168:
1133:
1129:
1119:
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1035:
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1010:
991:
985:
974:. Retrieved
944:(1): 51–61.
941:
937:
927:
919:the original
904:
885:
879:
869:
831:
794:
781:
761:
748:thermophilic
731:
704:
696:
664:
652:
640:
632:
623:multiplexing
617:
605:
585:
578:
566:
555:Calorimetric
543:
536:
527:Fluorescence
525:
522:Fluorometric
507:
505:
485:
484:
461:
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411:
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389:stopped-flow
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340:
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316:
314:
305:
296:
291:
287:
281:
258:
150:
91:
86:
84:
75:enzyme units
64:
61:Enzyme units
38:
37:
2159:Vertico SMI
2019:Protein NMR
1702:Hemosiderin
1583:Hyperplasia
1578:Hypertrophy
1554:Cell damage
1447:OpenWetWare
1242:Biophotonik
707:ionic bonds
637:Radiometric
568:Calorimetry
399:temperature
359:All enzyme
269:enzyme unit
2174:Categories
1868:Blood bank
1711:Lipofuscin
1707:Lipochrome
1683:Karyolysis
1612:Cell death
1593:Metaplasia
1457:2022-07-27
1328:10289/3552
1307:Biochem. J
1291:2019-07-02
1136:(7): 100.
976:2023-12-05
862:References
715:halophilic
596:luciferase
531:wavelength
512:hexokinase
496:hexokinase
478:forms. An
277:nanokatals
43:laboratory
2200:Pathology
1723:Steatosis
1666:Apoptosis
1603:Glandular
1588:Dysplasia
1520:Neoplasia
1515:Infection
1496:Pathology
821:MTT assay
784:substrate
457:MTT assay
321:substrate
131:×
47:enzymatic
1926:of study
1920:Proteins
1878:Serology
1673:Pyknosis
1619:Necrosis
1598:Squamous
1542:Ischemia
1413:11279227
1337:19849667
1285:Archived
1281:16002352
1219:17164337
1160:20981028
1111:18567619
1103:11554713
1068:31414611
970:Archived
966:40456258
958:16399643
840:See also
722:bacteria
711:proteins
476:oxidized
403:pressure
348:% purity
271:(U) = 1
2185:Enzymes
1924:methods
1751:Autopsy
1716:Melanin
1696:pigment
1573:Atrophy
1510:Disease
1372:9406427
1210:1750914
1187:Bibcode
1138:Bibcode
1059:6713963
751:archaea
735:protein
472:reduced
427:cuvette
284:gelatin
1922:: key
1411:
1370:
1335:
1279:
1217:
1207:
1158:
1109:
1101:
1066:
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998:
964:
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770:, and
549:lipase
498:using
455:. The
361:assays
309:enzyme
151:where
67:enzyme
2082:Assay
1525:Cause
1238:(PDF)
1107:S2CID
962:S2CID
801:rates
768:ionic
739:ionic
718:algae
468:NADPH
261:katal
71:molar
1409:PMID
1368:PMID
1333:PMID
1277:PMID
1215:PMID
1156:PMID
1099:PMID
1064:PMID
996:ISBN
954:PMID
832:para
803:and
741:and
466:and
464:NADH
346:The
339:The
273:μmol
53:and
41:are
1399:doi
1395:276
1360:doi
1356:118
1323:hdl
1315:doi
1311:425
1269:doi
1265:825
1205:PMC
1195:doi
1183:103
1146:doi
1091:doi
1087:296
1054:PMC
1044:doi
946:doi
942:329
890:doi
709:of
681:).
446:In
265:mol
2176::
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1177:.
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1105:.
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1085:.
1062:.
1052:.
1040:34
1038:.
1034:.
968:.
960:.
952:.
940:.
936:.
886:97
884:.
878:.
811:.
764:pH
724:.
657:.
551:.
518:.
401:,
279:.
89:.
77:.
57:.
1982:/
1968:/
1912:e
1905:t
1898:v
1709:/
1488:e
1481:t
1474:v
1460:.
1415:.
1401::
1374:.
1362::
1339:.
1325::
1317::
1294:.
1271::
1221:.
1197::
1189::
1162:.
1148::
1140::
1134:1
1113:.
1093::
1070:.
1046::
1004:.
979:.
948::
898:.
892::
502:.
242:V
218:r
193:t
188:n
163:a
135:V
127:r
123:=
118:t
113:n
108:=
104:a
20:)
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