SOURCE: Tritirachium album limber gene, recombinant
MOL. WEIGHT: 29.3 kDa
FORM: Solid: White powder
ACTIVITY: 1.07 kU/mg
UNIT DEFINITION: One unit is defined as the amount of protease that cleaves the FITC-labeled casein substrate to yield an amount of fluorescence equivalent to 1.0 nmol of unquenched FITC per minute at 37°C, pH 8.0
PURITY: Over 99% (Native PAGE and SDS PAGE assay), Molecular Biology Grade, RNase/DNase free
DNase ACTIVITY: None detectable enzyme activity with λ DNA after 6 hr incubation at 37°C
RNase ACTIVITY: None detectable ribonuclease activity after 16 hr incubation at 25°C
RECONSTITUTION: Reconstitute in 50 mM Tris-HCl (pH7.5), 3 mM CaCl2, 50% Glycerol
STORAGE CONDITIONS: Storage at 4°C or below is recommended (for solid enzyme). Aliquot the product after reconstitution to avoid repeated freeze/thaw cycles and store at -20°C. Also stable at room temperature for a year, if uncovered
EXPIRATION: Solid form for three years
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DESCRIPTION: Fabulously Active (FabActTM) Proteinase K is a highly reactive serine protease that displays the ability to digest native proteins, thereby inactivating enzymes such as DNase and RNase without recourse to a denaturation process. It is the most powerful proteinase among all proteinases characterized so far. It cleaves at the peptide bond adjacent to the carboxylic acid group of aliphatic, aromatic or hydrophobic amino acids.The application of the molecular grade FabActTM Proteinase K is similar to the Native Proteinase K and is used in the isolation or preparation of high molecular weight nucleic acids. However, FabActTM Proteinase K is highly pure and has a higher specific activity and is more stable at room temperature as compared to native Proteinase K. It is stable and active over a wide pH range of 4-12. It can be used on any situation to digest native and denatured proteins. FabActTM Proteinase K is also active with SDS, urea and EDTA and the most active temperature is 65°C.
Figure 1:Calculated Activity at 37°C, pH 8.0: Native Prot K: 802 nmol/min/mg (802 U/mg) or 16 kU/mL for 20 mg/mL solution; Molecular Grade Prot K: 1072 nmol/min/mg (1.07 kU/mg) or 21 kU/mL for 20 mg/mL solution. *One unit is defined as the amount of protease that cleaves the FITC-labeled casein substrate to yield an amount of fluorescence equivalent to 1.0 nmol of unquenched FITC per minute at 37°C, pH 8.0.
Figure 2: 1% Agarose-TBE gel lane description: Lane 1: 1 kB DNA Ladder; Lane 2: Whole Blood DNA isolated using Molecular Grade Pro K (total DNA yield from 500 μL blood sample = 3.6 μg; A260/280=1.90); Lane 3: Whole Blood DNA isolated using (Native Prot K) (total DNA yield from 500 μL blood sample = 2.9 μg; A260/280=1.89).
Removal of DNases and RNases when isolating DNA and RNA from tissues or cell lines for amplification reactions
INHIBITORS: The enzyme is inactivated by di-isopropyl fluorophosphates (DFP) and phenyl methane sulfonyl fluoride (PMSF)
HANDLING: Do not take internally. Wear gloves and mask when handling the product! Avoid contact by all modes of exposure