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Proteinase K-oppløsning, 1 ml

Proteinase K-oppløsning, 1 ml
€ 19,90
Inhalt: 1 Milliliter

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Leveringstid 2-5 virkedager

  • BS20.2551
Proteinase K-løsning (1 ml) 20 mg/ml; (i 50 mM Tris-HCl (pH 8,0), 1 mM CaCl2, 50 % glyserol).... mehr
Produktinformasjon

Proteinase K-løsning (1 ml)

20 mg/ml; (i 50 mM Tris-HCl (pH 8,0), 1 mM CaCl2, 50 % glyserol).

For pålitelig inaktivering av endogene nukleaser (DNaser, RNaser) under DNA- eller RNA-preparering. Proteinase K fra Bio&SELL er både svært aktiv (aktivitet 30 U/mg) og stabil og selvfølgelig fri for RNAser, DNAser og exonukleaser.

    Dens aktivitet kan inaktiveres av PMSF.
    Den anbefalte konsentrasjonen av proteinase K som skal brukes er 50 - 100 μg/ml for de fleste bruksområder.
    Kan brukes ved pH 4,3 - 12; optimal ved pH 8.

Weiterführende Links zu "Proteinase K-oppløsning, 1 ml"
Tekniske data

 

Kategorie: Serin-Endopeptidase (Hydrolase)

EC-Nummer: 3.4.21.64

Reaktionsart: Proteolytische Spaltung

Molekulargewicht: 28,9 kDa

Ursprungsorganismus: Tritirachium album

Expressionsorganismus: Komagataella phaffii (Pichia pastoris)

Form: Wässrige Lösung; 20 mg/ml

Reinheit: Proteingehalt > 70%
                  DNA-Gehalt ≤ 10 pg/mg; ≤ 200 pg/ml

Aktivität: ≥ 800 U/ml (1 U = 1 mAnsonU)

Ein U Proteinase K hydrolysiert Harnstoff-denaturiertes Hämoglobin und produziert das Farbequivalent von 1 µmol Tyrosin pro 1 min bei 37°C und pH 7.5 (Folin & Ciocalteu’s Methode)

Stabilität: pH 4,0-12,5 (pH-Optimum: 7,5 - 8,0); Auch in Anwesenheit von denaturierenden Agenzien (SDS, Harnstoff) stabil.
                   Ca
2+-Ionen (1-5 mM) schützen das Enzym vor Autolyse.

Aktivatoren: SDS, Harnstoff (Aktivierung durch Selbstverdau ist nicht nötig)

Inhibitoren: Hg2+-Ionen, PMFS, Phenol, DFP, Trypsin- bzw. Chymotrypsininhibitoren, Sulfhydryl-Reagenzien

Thermostabilität: aktiv bei 37-56°C (Optimale Aktiviät bei 50-55°C)

Inaktivierung: Hitzedenaturierung bei 75°C für 20 min

Lagerung: -20°C (aliquotiert), +4°C, RT

Solubilisierung: 20 mg/mL: Lösen in 50 ml destilliertem Wasser 
                              20–50 mg/mL: Lösen in definiertem Volumen in 50 mM Tris-HCl (pH 7,5 - 8,0), 1-5 mM Ca2+ (calcium chloride, calcium acetate)
                                                      oder 10 mM Tris-HCl (pH 7,5 - 8,0), 1-5 mM Ca2+ (calcium chloride, calcium acetate)

                              Für die für die Langzeitlagerung 50% Glycerol hinzufügen

Standardreaktionspuffer: Zelllyse: 50 mM Tris-HCl (pH 7,5), 5 mM CaCl2, 0,5% SDS
                                              DNA-Reinigung: 100 mM Tris-HCl (pH 8,0), 50 mM EDTA, 500 mM NaCl

Sicherheit: H315 + H319 + H334 + H317 + H335
                     P260 + P280 + P342 + P311

CAS-Nummer: 39450-01-6

 

FAQ

 

1. Bei welchen Anwendungen brauche ich Proteinase K?

  • Isolierung genomischer DNA aus Mausschwänzen:

                        Puffer: 20 mM Tris-HCl (pH 8,0), 400 mM NaCl, 5 mM EDTA (pH 8,0), 1% SDS
                        Konzentration: 400 µg/ml Protease K
                        Dauer:
Über Nacht
                        Temperatur:
55°C

  • Isolierung genomischer DNA aus kultivierten Zellen:

                        Puffer: 10 mM Tris-HCl (pH 8,0), 50 mM NaCl, 100 mM EDTA (pH 8,0), 0,5% SDS, 20 µg/ml DNase freie RNAse
                        Konzentration: 100 µg/ml Protease K
                        Dauer:
3h
                        Temperatur:
50°C

  • Schnelle Isolierung genomischer DNA (PCR-Template):

                        Puffer: 67 mM Tris-HCl (pH 8,8), 16,6 mM Ammoniumsulfat, 5 mM β-Mercaptoethanol, 6,7 mM MgCl2, 6,7 µM EDTA (pH 8,0), 1,7 µM SDS
                        Konzentration: 50 µg/ml Protease K
                        Dauer:
1h
                        Temperatur:
37°C

  • Reinigung von mRNA vor der cDNA-Synthese (Steigert oftmals die Ausbeute):

                        Puffer: 20 mM Tris-HCl (pH 7,5), 50 mM NaCl, 10 mM EDTA (pH 8,0), 0,1% SDS
                        Konzentration: 5 µg/ml Protease K
                        Dauer:
1-2h
                        Temperatur:
37°C

  • Aufreinigung von PCR-Ansätzen vor der Klonierung:

                        Ansatz: PCR-Ansätze + 20 mM Tris-HCl (pH 8,0), 400 mM NaCl, 5 mM EDTA (pH 8,0), 1% SDS
                        Konzentration: 400 µg/ml Protease K
                        Dauer:
über Nacht
                        Temperatur:
55°C

  • Aufreinigung von Plasmid-DNA vor in vitro Transkription:

                        Ansatz: Plasmid-DNA + 21 mM Tris-HCl (pH 8,0), 50 mM NaCl, 5 mM EDTA (pH 8,0), 0,5% SDS
                        Konzentration: 100 µg/ml Protease K
                        Dauer:
1h
                        Temperatur:
37°C

 

 

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