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COLLB-RO
Roche
Collagenase B
from Clostridium histolyticum
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About This Item
Polecane produkty
pochodzenie biologiczne
Clostridium histolyticum
Poziom jakości
sterylność
non-sterile
Formularz
lyophilized
opakowanie
pkg of 100 mg (11088807001)
pkg of 2.5 g (11088831001)
pkg of 500 mg (11088815001)
producent / nazwa handlowa
Roche
stężenie
0.5-2.5 mg/mL
optymalne pH
6.0-8.0
Zastosowanie
sample preparation
temp. przechowywania
2-8°C
Opis ogólny
Collagenase B is prepared from Clostridium histolyticum cultures by filtration, ammonium sulfate precipitation, dialysis, and lyophilization. Collagenase from Clostridium histolyticum is a collagenolytic enzyme. It interacts with collagen-like substances and is responsible for the degradation of collagen into small peptides. It is widely used for the disaggregation of many types of tissues (e.g., lung, heart, muscle, bone, adipose tissue, liver, kidney, cartilage, mammary gland, placentae, blood vessels, brain, tumors) and for the preparation of single-cell suspensions for the establishment of primary cell culture systems. Collagenase B is recommended when yield and viability are important.
Specyficzność
Collagenase degrades native collagen. Clostripain, trypsin-like enzymes, and neutral proteases degrade other proteins as well.
Zastosowanie
Clostridium collagenase from Roche has been used to prepare cells from many types of tissue, such as hepatocytes, adipocytes, pancreatic islets, epithelial cells, muscle cells, endothelial cells, etc. However, suitability of each lot of the enzyme for disruption of a particular tissue should be determined empirically.
Collagenase B has been used for preparation of cells from mouse aortas and colorectal cancer tissue.
Collagenase B has been used for preparation of cells from mouse aortas and colorectal cancer tissue.
Charakterystyka techniczna
Enzyme activity:
Collagenase activity: >0.15U/mg (according to Wünsch) (+25°C, 4-phenyl-azobenzyl-oxycarbonyl-Pro-Leu-Gly-Pro-D-Arg as the substrate)
Contaminating enzyme activities: trypsin, clostripain, and total proteolytic activity
Collagenase B has a normal to high collagenase activity and a higher than average clostripain activity (usually >10U/mg).
Inhibitors:
Collagenase inhibitors: EDTA, EGTA, Cys, His, DTT, 2-mercaptoethanol
Collagenase is not inhibited by serum.
Clostripain inhibitors: TLCK
Trypsin inhibitors: aprotinin, trypsin inhibitor (egg white, soybean), serum
Collagenase activity: >0.15U/mg (according to Wünsch) (+25°C, 4-phenyl-azobenzyl-oxycarbonyl-Pro-Leu-Gly-Pro-D-Arg as the substrate)
Contaminating enzyme activities: trypsin, clostripain, and total proteolytic activity
Collagenase B has a normal to high collagenase activity and a higher than average clostripain activity (usually >10U/mg).
Inhibitors:
Collagenase inhibitors: EDTA, EGTA, Cys, His, DTT, 2-mercaptoethanol
Collagenase is not inhibited by serum.
Clostripain inhibitors: TLCK
Trypsin inhibitors: aprotinin, trypsin inhibitor (egg white, soybean), serum
Definicja jednostki
Collagenase from Roche is assayed in Wünsch units (1 μmol of product formed per minute at +25 °C with Wünsch substrate).
Frequently, collagenase activities are given in Mandl units (1 μmol leucine liberated from collagen in 5 hours at +37 °C).
Unfortunately, there is no consistent conversion factor between the two units of activity, since the Mandl unit depends, in part, on the concentration of contaminating proteases in the collagenase preparation, an indefinable variable. A purer collagenase preparation would actually give a lower specific activity in Mandl units than a crude preparation. Clostridium preparations typically give conversion factors of approximately 1:1800 (e.g., a particular lot of Clostridium collagenase contained approximately 0.15 Wünsch U/mg and 250 Mandl U/mg).
Frequently, collagenase activities are given in Mandl units (1 μmol leucine liberated from collagen in 5 hours at +37 °C).
Unfortunately, there is no consistent conversion factor between the two units of activity, since the Mandl unit depends, in part, on the concentration of contaminating proteases in the collagenase preparation, an indefinable variable. A purer collagenase preparation would actually give a lower specific activity in Mandl units than a crude preparation. Clostridium preparations typically give conversion factors of approximately 1:1800 (e.g., a particular lot of Clostridium collagenase contained approximately 0.15 Wünsch U/mg and 250 Mandl U/mg).
Uwaga dotycząca przygotowania
Activator: Ca2+
Working concentration: 0.5 to 2.5 mg/ml
Storage conditions (working solution): -15 to -25 °C
Roche recommends reconstituting only the amount of lyophilizate needed for immediate use. The reconstituted solution can be stored at -15 to -25 °C for up to one week. Avoid repeated freezing and thawing since activity decreases after reconstitution.
Working concentration: 0.5 to 2.5 mg/ml
Storage conditions (working solution): -15 to -25 °C
Roche recommends reconstituting only the amount of lyophilizate needed for immediate use. The reconstituted solution can be stored at -15 to -25 °C for up to one week. Avoid repeated freezing and thawing since activity decreases after reconstitution.
Rekonstytucja
Reconstitution in any balanced salt solution (e.g., HBSS)
Inne uwagi
For life science research only. Not for use in diagnostic procedures.
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Hasło ostrzegawcze
Danger
Zwroty wskazujące rodzaj zagrożenia
Zwroty wskazujące środki ostrożności
Klasyfikacja zagrożeń
Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3
Organy docelowe
Respiratory system
Kod klasy składowania
11 - Combustible Solids
Klasa zagrożenia wodnego (WGK)
WGK 1
Temperatura zapłonu (°F)
does not flash
Temperatura zapłonu (°C)
does not flash
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Produkty
Enzyme Explorer Key Resource: Collagenase Guide.Collagenases, enzymes that break down the native collagen that holds animal tissues together, are made by a variety of microorganisms and by many different animal cells.
Powiązane treści
Kolagenazy, enzymy rozkładające natywny kolagen, który utrzymuje tkanki zwierzęce razem, są wytwarzane przez różne mikroorganizmy i wiele różnych komórek zwierzęcych.
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