Dapibus K mNGS (liquida)
Proteinase K est protease stabilis serine cum specificitate latis substratis.Multos servo in re publica depravat etiam coram detergents.Ex crystallo et hypothetica structurae testimonia enzyme indicat, ad subtilisin familiam cum activo catalytico trias (Asp.39-His69-Ser224).Praedominans situs bipertitis est vinculum peptidis adhaerens globi carboxyli amino acidi aliphatici et aromatici cum coetibus alpha amino inclusis.Vulgo usus est pro latis speciebus.Hoc proteinase K specialiter destinatum est pro mNGS.Comparatus cum altero proteinase K, etiam minus contaminationem acidum nucleicum cum eadem enzymatica operatione continet, quae melius applicationem amni mNGS curare potuit.
Repono Conditions
2-8℃ pro 2 annis
Specification
| Aspectus | Hyalina ut lux brunneis liquor |
| Actio | ≥800 U/ml |
| Dapibus Concentration | ≥20 mg/ml |
| Nickase | Nihil detectum |
| DNase | Nihil detectum |
| RNase | Nihil detectum |
Properties
| EC numerus | 3.4.21.64(Recombinant from Tritirachium album) |
| Isoelectric punctum | 7.81 |
| optimum pH | 7.0- 12.0 Fig |
| Optimum temperatus | 65 Fig |
| PH stabilitas | pH 4.5- 12.5 (25 , 16 h) Fig |
| Scelerisque status | Infra 50 (pH 8.0, 30 min) Fig |
| At stabilitas | Plus XC% actio pro XII mensis apud XXV ℃ |
| Activators | SDS, urea |
| Inhibitors | Diisopropyl fluorophosphate;phenylmethylsulfonyl fluoride |
Applications
1. Diagnostic ornamentum geneticae
2. RNA et DNA extraction kits
3. Extractio partium non-interdum e texturis, degradationum interdum immunditiarum, ut DNAvaccina et praeparatio heparin
4. Praeparatio chromosomatis DNA ab electrophoresi pulsante
5. Occidentis opprobrium
6. Enzymatica glycosylata albumin reagentia in vitro diagnosi
Cautiones
Gerunt chirothecas tutelares et goggles cum utendo vel appendendo, et post usum bene ventilatas.Productum hoc facere potest cutis reactionem allergic et gravem oculi irritationem.Si haustus, allergiam vel asthma signa vel dyspnoeam causare potest.Sit irritatio respiratorii causa.
Unitas definition
Una unitas (U) definitur quantum enzyme ad hydrolyze casein ad 1 μmol . producendum requiriturper- minutas tyrosinas sub his conditionibus.
Reagentia praeparatio
Reagens I: 1g casein lac in 50ml solutionis sodium phosphatae 0.1M resolutum (pH 8.0), incubatum 65-70 aqua per 15mina, ab aqua excitatum et dissolutum, refrigeratum, a sodium hydroxidum ad pH 8.0 accommodatum, et volumen certum. 100ml.
Reagens II: 0.1M Acidum trichloroaceticum, 0.2M Natrium acetatis, 0.3M Acidum aceticum.
Reagent III: 0.4M Na2CO3solutio.
Reagent IV: Forint iodi aqua pura dilutum 5 temporibus.
Reagens V: Enzyme diluens: 0.1M solutio phosphatae sodium (pH 8.0).
Reagens VI: solutio tyrosina: 0, 0.005, 0.025, 0.05, 0.075, 0.1, 0.25 umol/ml tyrosina cum 0.2 dissoluta.M HCl.
De modo procedendi
1. 0.5ml gerentis I praecalescentis ad 37℃, 0,5ml solutionis enzyme adde, bene misce et incubant.37℃ pro 10min.
2. Adde 1ml reagentes II ad resistendum reactionem, miscere bene, et pergere incubationem pro 30min.
3. solutio reactionis centrifugae.
4. Accipe 0.5ml supernatant, adde 2.5ml reagens III, 0.5ml reagens IV, misce bene et incubare 37℃pro 30mins.
5. OD660determinari ad OD1;blank control group: 0.5ml reagens V adhibetur reponere enzymesolution determinare OD660ut OD2, OD=OD1-OD2.
6. L-tyrosinum vexillum curvae: 0.5mL solutionem tyrosinam diversam intentionem L, 2.5mL Reagenti III, 0.5mL Reagenti IV in 5mL tubi centrifugii, incubare 37℃ pro 30min, deprehendere pro OD660ob diversam intentionem L-tyrosinam, tum curvae regulae Y=kX+b consecuta, ubi Y est L-tyrosina concentratio, X est OD.600.
Calculus
2: Totalis voluminis solutionis reactionis (ml)
0.5: Volume enzyme solutionis (mL)
0,5: Reactio liquida volumen in determinatione chromogenic (mL)
X: Reactio tempus (min)
Df: Dilutio multiplex
C: Enzyme concentration (mg/mL)
References
1. Wieger U & Hilz H. FEBS Lect.(1972);23:77.
2. Wieger U & Hilz H. Biochem.Biophys.Res.Communi.(1971);44:513.
3. Hilz, H.et al.EUR.J. Biochem.(1975);56: 103-108.
4. Sambrook Jet al. Molecular Cloning: A Laboratory Manual, 2nd edition, Frigidi Spring HarborLaboratorium Press, Frigidi Spring Harbour (1989).
Figurae
Fig.1 Optimum pH
100mM solutio quiddam: pH6.0-8.0, Na-phosphas;pH8.0-9.0, Tris-HCl;pH9.0-12.5, Glycine-NaOH.Enzyme concentration:1mg/mL
Fig.2 Optimum temperatus
Reactio in 20 mM K-phosphas quiddam pH 8.0.Enzyme retrahitur: 1 mg/mL
Fig.3 pH Stabilitas
25 , 16 h-tractatio cum 50 mM solutionis quiddam: pH 4.5- 5.5, acetate;pH 6.0-8.0, Na-phosphate;pH 8.0-9.0, Tris-.HCl.pH 9.0- 12.5, Glycine-NaOH.Enzyme retrahitur: 1 mg/mL
Fig.4 Scelerisque stabilitas
30 min-curatio cum 50 mM Tris-HCl quiddam, pH 8.0.Enzyme retrahitur: 1 mg/mL
Fig.5 Repono stabilisty at 25℃
