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P6556

Sigma-Aldrich

Proteinasa K from Tritirachium album

lyophilized powder, ≥30 units/mg protein

Sinónimos:

Endopeptidasa K

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About This Item

Número de CAS:
39450-01-6
Comisión internacional de enzimas:
3.4.21.64 (BRENDA, IUBMB)
Número CE:
254-457-8
Número MDL:
MFCD00132129
Código UNSPSC:
12352204
eCl@ss:
32160410
NACRES:
NA.54

formulario

lyophilized powder

Nivel de calidad

300

actividad específica

≥30 units/mg protein

mol peso

28.93 kDa

técnicas

DNA extraction: suitable

solubilidad

H2O: soluble 1 mg/mL, clear, colorless

actividad extraña

Dnase ≤30 Kunitz units/mg solid
RNase ≤0.003 Kunitz units/mg solid

Condiciones de envío

wet ice

temp. de almacenamiento

−20°C

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Aplicación

Útil para la inactivación proteolítica de nucleasas durante el aislamiento de ADN y ARN.
Elimina las endotoxinas que se unen a las proteínas catiónicas como la lisozima y la ribonucleasa A.
Se ha comunicado que es útil para el aislamiento de mitocondrias hepáticas, de levadura y de frijol mung
Determinación de la localización de las enzimas en las membranas
Tratamiento de cortes de tejido incluidos en parafina para exponer los sitios de unión de antígenos para el marcado con anticuerpos.
Digestión de proteínas de muestras de tejido cerebral para investigación de los priones en las encefalopatías espongiformes transmisibles (EET).
Product P6556 is provided as a lyophilized powder. Product P6556 has been used to break down human lens protein. Protease footprinting by Proteinase K digestion can reveal protein-protein surface interactions. The enzyme from Sigma has been used in the pre-hybridization step of chicken embryos. It has also been used for the enrichment of PrPSc, a prion protein that is present in sheep, hamster and mouse scrapie samples.
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 useful for the isolation of hepatic, yeast, and mung bean mitochondria
and is used to determine enzyme localization on membranes
It is used for the treatment of paraffin embedded tissue sections to expose antigen binding sites for antibody labeling and
for digestion of proteins from brain tissue samples for prions in Transmissible Spongiform Encephalopathies (TSE) research. Product P6556 is provided as a lyophilized powder. Product P6556 has been used to break down human lens protein.

Acciones bioquímicas o fisiológicas

Proteinase K 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 proteinasa K es una serín proteasa estable y muy reactiva. La evidencia procedente de estudios de la estructura cristalina y molecular indica que la enzima pertenece a la familia subtilisina con una tríada catalítica en el sitio activo (ASP39-HIS69-ser224). Es estable en una amplia variedad de entornos: pH, sales amortiguadoras, detergentes (SDS) y temperatura. En presencia de SDS al 0,1 - 0,5 %, la proteinasa K conserva su actividad y digerirá una variedad de proteínas y nucleasas en preparaciones de ADN sin comprometer la integridad del ADN aislado.

Definición de unidad

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).

también adquirido normalmente con este producto

Referencia del producto
Descripción
Precios

840857P

16:0-18:1 PA, powder

Pictogramas

Health hazardExclamation mark
GHS08,GHS07

Palabra de señalización

Danger

Frases de peligro

H315,H319,H334,H335

Clasificaciones de peligro

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

Órganos de actuación

Respiratory system

Código de clase de almacenamiento

11 - Combustible Solids

Clase de riesgo para el agua (WGK)

WGK 1

Punto de inflamabilidad (°F)

Not applicable

Punto de inflamabilidad (°C)

Not applicable

Equipo de protección personal

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

Certificados de análisis (COA)

Busque Certificados de análisis (COA) introduciendo el número de lote del producto. Los números de lote se encuentran en la etiqueta del producto después de las palabras «Lot» o «Batch»

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M M Kristjánsson et al.
European journal of biochemistry, 260(3), 752-760 (1999-04-02)
An extracellular serine proteinase purified from cultures of a psychrotrophic Vibrio species (strain PA-44) belongs to the proteinase K family of the superfamily of subtilisin-like proteinases. The enzyme is secreted as a 47-kDa protein, but under mild heat treatment (30
Rongzhu Cheng et al.
The Journal of biological chemistry, 279(44), 45441-45449 (2004-08-19)
We report here the isolation of a novel acid-labile yellow chromophore from the enzymatic digest of human lens proteins and the identification of its chemical structure by liquid chromatography-mass spectrometry, liquid chromatography-tandem mass spectrometry, and (1)H, (13)C, and two-dimensional NMR.
Nathaniel Denkers et al.
Developmental dynamics : an official publication of the American Association of Anatomists, 229(3), 651-657 (2004-03-03)
Multi-color whole-mount in situ hybridization is a powerful technique for comparing the spatial expression patterns of two or more genes in developing embryos. We have developed an amplified triple-label whole-mount fluorescence in situ hybridization (FISH) protocol that permits detection of
Michela Candelma et al.
Reproduction (Cambridge, England), 153(2), 123-132 (2016-11-03)
In vertebrates, the regulation of gametogenesis is under the control of gonadotropins (Gth), follicle-stimulating hormone (Fsh) and luteinizing hormone (Lh). In fish, the physiological role of Gths is not fully understood, especially in species with asynchronous ovarian development. To elucidate
Janus S Jakobsen et al.
Science advances, 5(7), eaaw4304-eaaw4304 (2019-07-17)
The key myeloid transcription factor (TF), CEBPA, is frequently mutated in acute myeloid leukemia (AML), but the direct molecular effects of this leukemic driver mutation remain elusive. To investigate CEBPA mutant AML, we performed microscale, in vivo chromatin immunoprecipitation sequencing
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Artículos

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).

Protocolos

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).

RNAi In Situ

In Situ Hybridization of Whole-Mount Mouse Embryos with RNA Probes: Hybridization, Washes, and Histochemistry

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