Proteinase k has a broad temperature profile, retaining 80% of its activity at tempera. The proteinase k based solution is designed to break the protein crosslinks, therefore unmask the antigens and epitopes in. Proteinase k safety data sheet according to federal register vol. Purchaser must determine the suitability of the products for their. Proteinase definition of proteinase by the free dictionary. Next the polysomes are dissociated with sds and proteinase k and finally the protein is removed by several phenolcholoroform extractions 1, 2. Proteinase k was isolated from a fungus able to grow. It also degrades any nucleases andor enzymes that may be present in the sample. Contamination of a relatively rnasefree cytoplasmic environment with nuclear nucleases is thus minimised.
Reconstitute proteinase k dissolve the proteinase k. Proteinase k solution catalog number am2546, am2548 pub. For protease digestion during dna and rna preparation. Prepare a stock solution in the vial by adding 2 ml of a solution of 0. Proteinase k prok is a serine endopeptidase with a broad spectrum of action, isolated from the filtrate of the fungus tritirachium album limber characteristics of proteinase k from tritirachium album limber molecular weight. Thermo scientific proteinase k is a broadrange endolytic protease widely used for digestion of proteins in nucleic acid preparations. It possesses a high specific activity which remains stable over a wide range of temperatures and ph values with substantially increased activity at. Qiagen protease is completely free of dnase and rnase activities. An endopeptidase, such as pepsin, trypsin, or papain.
It is particularly suitable for short digestion times. Proteinase k prepared at 20mgml in 10mm trishcl, 1mm calcium acetate, ph 7. Sigma brand products are sold through sigmaaldrich, inc. Protease k, endopeptidase k, tritirachium alkaline proteinase, molecular biology grade proteinase k. This is of vital importance since these chemical comp. Proteinase k is active in a wide range of buffers including all neb specific restriction endonuclease buffers. The chromatographically purified 46kda proteinase kresistant protein of mycoplasma orale was an arginine deiminase. Proteinase k fungal, 40 unitsmg1 100 mg 1 g 2c8c 1 one manson unit is described as that amount of enzyme that liberates1. Proteinase k from tritirachium album lyophilized powder, bioultra. Proteinase k is a stable and highly reactive serine protease.
Identification and characterization of proteinase k. In an attempt to define the nature of the scrapie agent prusiner used a variety of hydrolytic enzymes such as proteinase k pk to assess its grade of infectivity after inactivation. P480 proteinase k is a nonspecific serine protease often used in the preparation of genomic dna and can react with our without sds, edta and urea. The buffering conditions used ensure that the enzyme retains its full activity and offers convenience to the researchers.
The enzyme was discovered in 1974 in extracts of the fungus engyodontium album. Proteases catalyze the breakdown of contaminating proteins present in the solution to its component amino acids. Raising the temperature of the reaction from 37c to 50c 60c can increase the activity several folds. In molecular biology proteinase k is a broadspectrum serine protease. Proteinase ka subtilisin related enzyme with disulfide bonds. Qiagen proteinase k is a subtilisintype protease, which cleaves at the carboxyl side of hydrophobic, aliphatic and aromatic amino acids. Proteinase k is widely used in the purification of native rna and dna from tissues or cell lines, as well as for many other applications. Proteinase k is not inhibited by edta, iodoacetic acid, tlck or tpck. Proteinase k is also active in chelating agents such as edta 4 and activity is stimulated in up to 2% sds or 4 m urea 3. Proteinase k is a proteolytic enzyme a serine protease that is purified from the mold tritirachium album. The following are examples illustrating this application.
Proteinase k was isolated from a fungus able to grow on keratin and the. Us state regulations the product andor components does not appear on any state right to know lists or. Proteinase k cleaves peptide bonds at the carboxylic sides of aliphatic, aromatic, or hydrophobic amino acid. This product is guaranteed for 6 months from the date of receipt, if properly stored. Proteinase k is a highly active 28,904da serine protease isolated from the fungus tritirachium album 1.
Proteinase k, solution a concentrated, ready to use liquid formulation. The recommended working concentration of proteinase k is 0. Proteinase k, recombinant, 20 mgml solution, molecular. Enzymatic reaction image will open in a new window. The enzyme belongs to the group of subtilisinerelated serine proteases and is strongly inhibited by pmsf. Prepare a stock solution in the vial by adding 2 ml. It cleaves peptide bonds adjacent to the carboxylic group of aliphatic and aromatic amino acids and is useful for general digestion of protein in biological samples.
The proteinase k, lyophilized powder can be prepared as a 20mgml stock solution in water and stored in aliquots at 20c the enzyme is also available as a readytouse concentrated stock solution 600mauml that is convenient for routine use in most applications. An interesting characteristic of proteinase k is that it retains its activity in the presence of sodium dodecyl sulphate sds or urea. Longlife proteinase k is stable for a year if stored correctly. What is the difference between qiagen protease and qiagen. Proteinase k is able to digest hair, hence, the name. The enzyme was discovered in 1974 in extracts of the fungus engyodontium album formerly tritirachium album. Proteinase k, recombinant, pcr grade solution from pichia pastoris. Louis, mo were added to ampli fication reactions alone and in combina tion. C contents quantity storage conditions proteinase k, 20 mgml am2546. It possesses a high specific activity over a wide range of temperatures and ph values with substantially increased activity at higher temperature. The smallest peptide to be hydrolysed is a tetrapeptide. Proteinase k is a highly active serine protease with broad cleavage specificity on native and denatured proteins. Full text get a printable copy pdf file of the complete article 1. Crystal structure of calciumfree proteinase k at 1.
The enzyme has abroad specificity, cleaving peptide bonds cterminal to a number of amino acids. Many proteases have proven useful for monitoring translocation in this fashion including protease k or trypsin. Proteinase k is a very stable protease, active in wide ph and temperature ranges. It possesses a high specific activity which remains stable over a wide range of temperatures and ph values with substantially increased activity at higher temperature. The activity of proteinase k is increased in the presence of denaturants such as sds 1% and at elevated temperature 5060c. Proteinase k is not inactivated by chelating reagents such as edta or detergents such as sds and is active over a wide range of ph 412. Proteinase k solution, rna grade thermo fisher scientific. Proteinase k 20 mgml purchase as a lyophilized powder and dissolve at a concentration of 20 mgml in sterile 50 mm tris ph 8.
May 26, 2010 many proteases have proven useful for monitoring translocation in this fashion including protease k or trypsin. Proteinase k is a subtilisintype protease isolated from the saprophytic fungus tritirachiumalbum and is particularly suitable for short digestion times. Proteinase k solution 10ml 20mgml proteinase k solution. Proteinase k 20 mgml solution is a nonspecific serine protease often used in the preparation of genomic dna and can react with our without sds, edta and urea. Proteinase k is commonly used in molecular biology to digest protein and remove contamination from preparations of nucleic acid. Proteinase k solution 5ml 20mgml genomic pcr grade proteinase k solution. It is highly suited to this application since the enzyme is active. Proteinase k antigen retrieval protocol ihc world, llc. A simple universal dna extraction procedure using sds and. Rna extraction by the proteinase k method springerlink. Evidence from crystal and molecular structure studies indicates the enzyme belongs to the subtilisin family with an activesite catalytic triad asp 39 his 69 ser 224.
The enzyme belongs to serine proteases and cleaves peptide bonds following aliphatic, aromatic or hydrophobic amino acids. Proteinase k solution 20 mgml thermo fisher scientific. It degrades proteins even in the presence of detergents. Qiagen protease and qiagen proteinase k offer broad substrate specificity with high activity for a wide range. Stimulation of proteinase k action by denaturing agents. P2308 sigmaaldrich proteinase k from tritirachium album lyophilized powder, bioultra. Proteinase k is a very active and highly stable endopeptidase. Proteinase k, produced by the fungus tritirachium album limber, is a serine protease that exhibits a very broad cleavage specificity. Proteinase k products are free of detectable dnase and rnase. Proteinase k, recombinant, 20 mgml solution, molecular grade. Material safety data sheet msds for proteinase k solution.
The recommended working concentration is 50100 gml for protein removal and enzyme inactivation and up to 2 mgml for tissue treatment. Addition of proteinase k to nucleic acid preparations rapidly inactivates nucleases that might otherwise degrade the dna or rna during purification. The vial of proteinase k from sigma protease, proteinase k type xi, cat. Proteinase k is a subtilisinrelated serine protease that hydrolyzes a variety of peptide bonds and is used to cleanup enzymatic reactions or lysates. The smallest peptide to be hydrolysed is a tetrapeptide 1. Each aliquot can be thawed and refrozen several times but should then be discarded. Proteinase k is a nonspecific, subtilisinrelated serine protease with a very high specific activity.
Milliporesigma proteinase k solution, 600mauml life. Formalin or other aldehyde fixation forms protein crosslinks that mask the antigenic sites in tissue specimens, thereby giving weak or false negative staining for immunohistochemical detection of certain proteins. Recently i bought 100 mg proteinase k power from roche 3115879001, proteinase k,recomb. Isolated from the fungus engyodontium album formerly tritirachium album, proteinase k exhibits broad substrate specificity and degrades many proteins in the native state, even in the presence of detergents. Reagents designed and manufactured under cgmp controls suitable for use in an ivd application. Recombinant proteinase k dissolved in water to a final concentration of 20 mgml.
Proteinase k is useful in purifying high molecular weight genomic nucleic acids from cells and. Proteinase k is an endolytic protease that cleaves peptide bonds at the carboxylic sides of aliphatic, aromatic or hydrophobic amino acids. The enzyme is active in the presence of 1% sds, 1% triton x100, 4m urea, 3m guanidine hcl or 5% tween20. How to prevent complete degradation of tissue by proteinase k. Proteinase k is inhibited by diisopropyl fluorophosphate dfip, and phenylmethanesulfonyl fluoride pmsf. Peptidase k, endoproteinase k, endopeptidase k temperature prof product description proteinase k is a stable serine protease with broad substrate specificity.
Proteinase k or solution is a highly active and stable protease with low cutting specificity. Proteinase k is a serine protease and the main proteolytic enzyme produced by the fungus tritirachium album limber. Also, pyrogenfree water was used that had been shown to be free of con taminating bacterial dna. Proteinase k from the fungus engyodontium album is a nonspecific serine protease that is useful for general digestion of proteins. Identification and characterization of proteinase kresistant. Proteinase k from engyodontium album formerly tritirachium album for molecular biology catalog number p2308 storage temperature 20 c cas rn 39450016 e.
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