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P8250

Sigma-Aldrich

Perossidasi

Type II, essentially salt-free, lyophilized powder, 150-250 units/mg solid (using pyrogallol)

Sinonimo/i:

Donatore: idrogeno-perossido ossidoreduttasi, Perossidasi di rafano

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About This Item

Numero CAS:
9003-99-0
Classificazione EC (Enzyme Commission):
1.11.1.7 (BRENDA, IUBMB)
Numero CE:
232-668-6
Numero MDL:
MFCD00071339
Codice UNSPSC:
12352204
eCl@ss:
32160410
NACRES:
NA.54

Origine biologica

horseradish

Livello qualitativo

300

Tipo

Type II

Forma fisica

essentially salt-free, lyophilized powder

Attività specifica

150-250 units/mg solid (using pyrogallol)

PM

~44 kDa

Solubilità

0.1 M phosphate buffer: soluble (pH 6.0)
H2O: soluble

Rapporto di assorbanza

RZ ≥1.8

applicazioni

diagnostic assay manufacturing

Temperatura di conservazione

2-8°C

InChI

1S/H2O3/c1-3-2/h1-2H
JSPLKZUTYZBBKA-UHFFFAOYSA-N

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Descrizione generale

La perossidasi di rafano si isola dalle radici del rafano (Amoracia rusticana) e appartiene al gruppo delle perossidasi con ferroprotoporfirina. L′HRP è un polipeptide a singola catena che contiene quattro ponti disolfuro. È una glicoproteina che contiene il 18% di carboidrati. La composizione dei carboidrati include galattosio, arabinosio, xilosio, fucosio, mannosio, mannosamina e galattosamina, a seconda del singolo isozima. Il peso molecolare (circa 44 kDa) include la catena polipeptidica (33.890 Dalton), la emina con Ca2+ (circa 700 Dalton) e il carboidrato (circa 9.400 Dalton). Esistono almeno sette isoenzimi di HRP. Il punto isoelettrico degli isoenzimi della perossidasi di rafano è compreso fra 3,0 e 9,0.

Applicazioni

Horseradish peroxidase (HRP) is isolated from horseradish roots (Amoracia rusticana). It is used in biochemistry applications such as western blots, ELISA and Immunohistochemistry. Horseradish peroxidase is used to amplify a weak signal and increase detectability of a target molecule, such as a protein. Horseradish peroxidase, product P8250, has been used to study nonoral antigens in inflamed gingiva and Ebola virus glycoprotein toxicity.
The enzyme has been used as a comparison during the peroxidase assay of extract from mature tall fescue leaf blades. It has also been used to measure H2O2 production.

Azioni biochim/fisiol

HRP readily combines with hydrogen peroxide (H2O2) and the resultant [HRP-H2O2] complex can oxidize a wide variety of hydrogen donors. The optimal pH is 6.0-6.5 and the enzyme is most stable in the pH range of 5.0-9.0. HRP can be conjugated to antibodies by several different methods including glutaraldehyde, periodate oxidation, through disulfide bonds, and also via amino and thiol directed cross-linkers. It is smaller and more stable than the enzyme labels β-galactosidase and alkaline phosphatase. Hence, it is the most desired label. Also, its glycosylation leads to lower non-specific binding. It is also used for the determination of glucose and peroxides in solution. Sodium azide, cyanide, L-cystine, dichromate, ethylenethiourea, hydroxylamine, sulfide, vanadate, p-aminobenzoic acid, and Cd2+, Co2+, Cu2+, Fe3+, Mn2+, Ni2+, Pb2+ ions are found to inhibit the enzyme activity.
When incubated with a substrate, horseradish peroxidase produces a coloured, fluorimetric, or luminescent derivative of the labeled molecule, allowing quantification. Horseradish peroxidase has been shown to slightly reduce the level of inhibition in a cydAB mutant.

Qualità

Preliminary work shows it to contain at least five isoenzymes.

Definizione di unità

Una unità di pirogallolo forma 1,0 mg di purpurogallina da pirogallolo in 20 secondi a pH 6,0 e 20 °C.

Risultati analitici

Il RZ (Reinheitszahl) è il rapporto di assorbanza A403/A275 determinato a 0,5-1,0 mg/mL in acqua deionizzata. Misura il contenuto di emina, non l′attività enzimatica. Anche preparazioni con un RZ elevato possono avere una bassa attività enzimatica.

Altre note

Per maggiori informazioni sulla perossidasi visitare la pagina www.sigma-aldrich.com/enzymeexplorer.

Inibitore

N° Catalogo
Descrizione
Determinazione del prezzo

Substrato

N° Catalogo
Descrizione
Determinazione del prezzo

Pittogrammi

Health hazard
GHS08

Avvertenze

Danger

Indicazioni di pericolo

H334

Consigli di prudenza

P261 - P284 - P501

Classi di pericolo

Resp. Sens. 1

Codice della classe di stoccaggio

13 - Non Combustible Solids

Classe di pericolosità dell'acqua (WGK)

WGK 1

Dispositivi di protezione individuale

Eyeshields, Gloves, type N95 (US)

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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Peroxidase

Peroxidase (POD) EIA grade, from horseradish

Roche

10814407001

Peroxidase (POD)

W E Bullock et al.
The Journal of experimental medicine, 165(1), 195-210 (1987-01-01)
The principal host cell of H. capsulatum (Hc) is the M phi within which the pathogenic yeast phase of the fungus multiplies during active disease. The initial interaction between Hc yeasts and M phi therefore is a crucial step in
S M Mallison et al.
Infection and immunity, 56(4), 823-830 (1988-04-01)
In vitro experimentation indicates that periodontitis-associated bacteria contain potent polyclonal B-cell activators (PBA). We reasoned that if PBA were operative in vivo, plasma cells specific for nonoral antigens should be present in the inflamed gingival tissues, which are characterized by
J W Macadam et al.
Plant physiology, 99(3), 872-878 (1992-07-01)
Cessation of cell expansion has been associated with cell wall cross-linking reactions catalyzed by peroxidase. This study utilized two genotypes of tall fescue (Festuca arundinacea Schreb.) that differ in length of the leaf elongation zone to investigate the relationship between
Chuan Chiang-Ni et al.
Journal of microbiology, immunology, and infection = Wei mian yu gan ran za zhi, 49(6), 837-842 (2015-02-05)
The phenotypic heterogeneity of the human pathogen Streptococcus pyogenes [group A streptococcus (GAS)] is associated with bacterial virulence variation. During invasive GAS infection, mutations in the two-component regulatory system covR/covS leads to increases in hyaluronic acid capsule production, virulence genes
Nancy J Sullivan et al.
Journal of virology, 79(1), 547-553 (2004-12-15)
Ebola virus infection causes a highly lethal hemorrhagic fever syndrome associated with profound immunosuppression through its ability to induce widespread inflammation and cellular damage. Though GP, the viral envelope glycoprotein, mediates many of these effects, the molecular events that underlie
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