Proteinase K (pulveris Lyophilized)
Cat No: HC4500A
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 pro lataproprietatem.
Repono Conditions
2-8℃ breve tempus repositionis, 25~-15℃ longum tempus repositum.Arida pulveris status -25~-15℃ valet pro 3 annis;pulvis enzyme in apto volumine dissolvatur.
Specification
| Aspectus | Alba ut off-album amorpho lyophilized pulveris |
| Actio | ≥30 U/mg |
| DNase | Nihil detectum |
| RNase | Nihil detectum |
Properties
| EC numerus | 3.4.21.64 (Recombinant from Tritirachium album) |
| M. pondus | 29 kDa (SDS-PAGE) |
| Isoelectric punctum | 7.81 |
| optimum pH | 7.0-12.0 Fig.1 |
| Optimum temperatus | 65 Fig.2 |
| PH stabilitas | pH 4.5-12.5 (25℃, 16h) Fig |
| Scelerisque status | Infra 50℃ (pH 8.0, 30min) Fig |
| 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 textuum, degradatione immunditiarum interdum, ut DNA vaccina 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.
Assay
Unitas definition
Unitas una (U) definitur quantum enzyme ad hydrolyzi casein requiritur ad 1 μmol tyrosinum per minutum producendum 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 dissoluta cum 0.2M HCL.
De modo procedendi
1. 0,5ml reagentia I prae-calefactum ad 37℃, 0,5ml solutionis enzyme adde, bene misce, et incubant 37℃ pro 10min.
2. Adde 1 ml reagentes II ad resistendum reactionem, miscere bene, et pergere incubationem pro 30min.
3. solutio reactionis centrifugae.
4. Accipe 0.5ml supernatantem, adde 2.5ml reagens III, 0.5ml reagens IV, misce bene et incubare 37℃ pro 30min.
5. OD660determinari ad OD1;blank control group: 0.5ml reagens V adhibetur ad solutionem enzyme reponere ad 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 OD.660ob diversam intentionem L-tyrosinam, tum curvae regulae Y=kX+b consecuta, ubi Y est concentratio L- tyrosina, 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);
4. Sambrook Jet al. Molecular Cloning: A Laboratory Manuale, 2nd edition, Frigidum Spring HarborLaboratory Press, Frigidum Spring Harbour (1989).
Fig. 2 Optimum temperatus
Reactio in 20 mM K-phosphas quiddam pH 8.0.Enzyme concentration: 1mg/mL
Fig Stabilitas
25℃, 16 h-tractatio cum 50mM 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 concentration: 1mg/mL
Fig stabilitas
30 min-tractatio cum 50mM Tris-HCL quiddam, pH 8.0.Enzyme concentration: 1mg/mL
Fig stabilisty at 25℃
