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FOR RESEARCH USE ONLY! 05/16 FabActTM Proteinase K (recombinant), Molecular Grade (Catalog # 9213-5, -25; 20 mg/mL Solution) ALTERNATE NAMES: Protease K, Endopeptidase K, Tritirachium alkaline proteinase CATALOG NO.: 9213-5, -25 AMOUNT: 5 ml, 25 ml SOURCE: Tritirachium album limber gene, recombinant MOL. WEIGHT: 29.3 kDa FORM: Colorless liquid 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: Reconstituted in 50 mM Tris-HCl (pH 7.5), 3 mM CaCl2, 50% Glycerol STORAGE CONDITIONS: Aliquot the product to avoid repeated freeze/thaw cycles and store at -20°C EXPIRATION: Liquid form for one year 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. 1 2 3 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). 155 S. Milpitas Blvd., Milpitas, CA 95035 USA | T: (408)493-1800 F: (408)493-1801 | www.biovision.com | [email protected] FOR RESEARCH USE ONLY! APPLICATIONS: 05/16 Removal of DNases and RNases when isolating DNA and RNA from tissues or cell lines for amplification reactions Isolation of mRNA or genomic DNA from different tissues including mouse tail or from cultured cells For modifying glycoprotein for structural studies To treat tissue sections for in situ hybridization Improving cloning efficiency of PCR products For isolating bio-product to remove protein contaminants in industries such as leather, food, feed, medicine intermediates, etc 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 RELATED PRODUCTS: FabActTM Proteinase K (recombinant), Molecular Grade (Solid) (Cat. No. 9212-100, -500, -1G, -10G) Binuclease (Solid) (Cat. No. 9214-10, -50, -100, -500) Binuclease (Liquid) (Cat. No. 9215-20, -200) Protease Activity Assay kit (Cat. No. K781-100) Protease Inhibitor Cocktail (Cat. No. K271-500) EZBlock Protease Inhibitor Cocktail, EDTA-Free (Cat. No. K272-1, -5; K272-set) EZBlockTM Protease Inhibitor Cocktail II (Cat. No. K277-set) EZBlockTM Protease Inhibitor Cocktail III (Cat. No. K278-set) EZBlockTM Protease Inhibitor Cocktail IV (Cat. No. K279-1; K279-set) EZBlockTM Universal Protease and Phosphatase Inhibitor Cocktail (Cat. No. K283-1; K283-set) EZBlockTM Universal Protease and Phosphatase Inhibitor Cocktail, EDTA-Free (Cat. No. K284-1; K284-set) Protease and Phosphatase Inhibitor Cocktails, Mini Brochure (Cat# BVBPPIC-2011) FOR RESEARCH USE ONLY! Not to be used on humans. 155 S. Milpitas Blvd., Milpitas, CA 95035 USA | T: (408)493-1800 F: (408)493-1801 | www.biovision.com | [email protected]