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P8044

Sigma-Aldrich

Proteinasi K

≥3.0 unit/mg solid, lyophilized powder

Sinonimo/i:

Endopeptidasi K

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About This Item

Numero CAS:
39450-01-6
Classificazione EC (Enzyme Commission):
3.4.21.64 (BRENDA, IUBMB)
Numero CE:
254-457-8
Numero MDL:
MFCD00132129
Codice UNSPSC:
12352204
eCl@ss:
32160410
NACRES:
NA.54

Origine biologica

maize
microbial (T.album
T. ALBUM)
plant seeds (barley)
soybean
yeast

Livello qualitativo

200

Forma fisica

lyophilized powder

Attività specifica

≥3.0 unit/mg solid

PM

28.93 kDa

Composizione

Protein, ≥15% biuret

tecniche

DNA extraction: suitable

applicazioni

diagnostic assay manufacturing

Temperatura di conservazione

−20°C

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Categorie correlate

Applicazioni

Utile per l′inattivazione proteolitica delle nucleasi durante l′isolamento di DNA ed RNA.
Rimuove le endotossine che si legano alle proteine cationiche come lisozima e ribonucleasi A.
Segnalato come utile per l′isolamento dei mitocondri epatici, di lievito e di fagiolo mungo
Determinazione della localizzazione degli enzimi sulle membrane
Trattamento di sezioni di tessuto incluse in paraffina per esporre i siti di legame dell′antigene per la marcatura dell′anticorpo.
Digestione delle proteine da campioni di tessuto cerebrale per la ricerca sui prioni nell′encefalopatia spongiforme trasmissibile (EST).
Proteinase K is useful for the proteolytic inactivation of nucleases during the isolation of DNA and RNA. It is used for the removal of endotoxins bound to cationic proteins such as lysozyme and ribonuclease A. It is also useful for the isolation of hepatic, yeast, and mung bean mitochondria and is used to determine enzyme localization on membranes. Furthermore, it is used for the treatment of paraffin embedded tissue sections to expose antigen binding sites and for digestion of proteins from brain tissue samples. Product P8044 is provided as a lyophilized powder.
The enzyme from Sigma has been used to degrade complex I (NADH:ubiquinone oxidoreductase) prior to extraction of ubiquinone from Yarrowia lipolytica. It has been used to examine the effect of Proteinase k on Pythium ultimum. It has also been used to maximize the variety of peptide bonds hydrolyzed in the sediment slurry without autolytic production of amino acids from the enzyme itself.

Azioni biochim/fisiol

Proteinase K is highly active towards native proteins. It has a broad specificity and degrades many proteins even in the native state. It mainly cleaves the peptide bond adjacent to the carboxyl group of aliphatic and aromatic amino acids with blocked α-amino groups. The optimum pH is between 7.5-9.0 and the isoelectric point is 8.9. Ca2+ (1-5 mM) is required for activation. Proteinase K is inhibited by diisopropyl fluorophosphate (DFIP), and phenylmethanesulfonyl fluoride (PMSF).
La proteinasi K è una serin proteasi stabile e molto reattiva. Le evidenze emerse da studi sulla struttura cristallina e molecolare indicano che l′enzima appartiene alla famiglia delle subtilisine con una triade catalitica nel sito attivo (Asp39-His69-Ser224). È stabile in un vasto assortimento di ambienti: pH, sali tampone, detergenti (SDS) e temperatura. In presenza di SDS 0,1-0,5%, la proteinasi K conserva la sua attività e digerisce una serie di proteine e nucleasi nelle preparazioni di DNA senza compromettere l′integrità del DNA isolato.

Definizione di unità

One unit will hydrolyze urea-denatured hemoglobin to produce color equivalent to 1.0 μmole of tyrosine per min at pH 7.5 at 37 °C (color by Folin-Ciocalteu reagent).

Pittogrammi

Health hazardExclamation mark
GHS08,GHS07

Avvertenze

Danger

Indicazioni di pericolo

H315,H319,H334,H335

Classi di pericolo

Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3

Organi bersaglio

Respiratory system

Codice della classe di stoccaggio

11 - Combustible Solids

Classe di pericolosità dell'acqua (WGK)

WGK 1

Punto d’infiammabilità (°F)

Not applicable

Punto d’infiammabilità (°C)

Not applicable

Dispositivi di protezione individuale

dust mask type N95 (US), Eyeshields, Faceshields, Gloves

Certificati d'analisi (COA)

Cerca il Certificati d'analisi (COA) digitando il numero di lotto/batch corrispondente. I numeri di lotto o di batch sono stampati sull'etichetta dei prodotti dopo la parola ‘Lotto’ o ‘Batch’.

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Millipore

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Proteinasi K

Proteinasi K, ricombinante, grado PCR Solution from Pichia pastoris

Roche

RPROTKSOL-RO

Proteinasi K, ricombinante, grado PCR

Proteinasi K, ricombinante, grado PCR Lyophilizate from Pichia pastoris

Roche

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Proteinasi K, ricombinante, grado PCR

P Bermejo-Alvarez et al.
Biology of reproduction, 79(4), 594-597 (2008-06-27)
It has been reported that the mammalian female could have a preconceptual influence on the sex of her offspring, and it has been hypothesized that this influence could go some way toward accounting for the reported lower fertility following insemination
C Dunne et al.
Microbiology (Reading, England), 146 ( Pt 8), 2069-2078 (2000-08-10)
Stenotrophomonas maltophilia W81 can protect sugar beet against PYTHIUM:-mediated damping-off disease through the production of an extracellular protease. Here, the proteolytic enzyme of W81 was purified by anion-exchange chromatography and characterized as a serine protease. The purified enzyme was fungicidal
Bioavailable amino acids in sediments: A biomimetic, kinetics-based approach
Mayer LM, et al.
Limnology and Oceanography, 40(3), 511-520 (1995)
Stefan Dröse et al.
Biochimica et biophysica acta, 1556(1), 65-72 (2002-09-28)
NADH:ubiquinone oxidoreductase (complex I) is the largest multiprotein complex of the mitochondrial respiratory chain. His-tagged complex I purified from the strictly aerobic yeast Yarrowia lipolytica exhibited electron transfer rates from NADH to n-decylubiquinone of less than 2% when compared to
Enzymes of Molecular Biology
M.M. Burrell
Methods in Molecular Biology, 16, 307-307 (1993)
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Articoli

Enzymatic Assay of Proteinase K with Hemoglobin Substrate

Proteinase K (EC 3.4.21.64) activity can be measured spectrophotometrically using hemoglobin as the substrate. Proteinase K hydrolyzes hemoglobin denatured with urea, and liberates Folin-postive amino acids and peptides. One unit will hydrolyze hemoglobin to produce color equivalent to 1.0 μmol of tyrosine per minute at pH 7.5 at 37 °C (color by Folin & Ciocalteu's Phenol Reagent).

Enzymatic Assay of Proteinase K with Hemoglobin Substrate

Proteinase K (EC 3.4.21.64) activity can be measured spectrophotometrically using hemoglobin as the substrate. Proteinase K hydrolyzes hemoglobin denatured with urea, and liberates Folin-postive amino acids and peptides. One unit will hydrolyze hemoglobin to produce color equivalent to 1.0 μmol of tyrosine per minute at pH 7.5 at 37 °C (color by Folin & Ciocalteu's Phenol Reagent).

Protocolli

Enzymatic Assay of Proteinase K (EC 3.4.21.64)

Proteinase K (EC 3.4.21.64) activity can be measured spectrophotometrically using hemoglobin as the substrate. Proteinase K hydrolyzes hemoglobin denatured with urea, and liberates Folin-postive amino acids and peptides. One unit will hydrolyze hemoglobin to produce color equivalent to 1.0 μmol of tyrosine per minute at pH 7.5 at 37 °C (color by Folin & Ciocalteu's Phenol Reagent).

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