Proteinase K Powder
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Invitrogen™

Proteinase K Powder

Proteinase K from the fungus Engyodontium album is a non-specific serine protease that is useful for general digestion of proteins.ProteinaseRead more
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Catalog NumberQuantity
AM2542100 mg
AM2544500 mg
Catalog number AM2542
Price (MXN)
-
Quantity:
100 mg

Proteinase K from the fungus Engyodontium album is a non-specific serine protease that is useful for general digestion of proteins.

Proteinase K Remains Active:
• Over a wide pH range (optimal activity between 6.5 and 9.5)
• Under denaturing conditions (e.g. the presence of SDS or urea)
• In the presence of metal chelating agents (e.g. EDTA)
• At comparatively high temperatures (optimum digestion temperature is 65°C)

Applications: Removal of endogenous nucleases during the preparation of DNA and RNA; preparation of tissue sections for in situ hybridization.

Performance and Quality Testing: Endodeoxyribonuclease and exodeoxyribonuclease assays; liquid form tested for absence of RNase activity.

Unit Definition: One mAnson unit is described as that amount of enzyme that liberates 1 μmole of Folin-positive amino acid within one minute at 37°C using hemoglobin as a substrate.

For Research Use Only. Not for use in diagnostic procedures.
Specifications
Host SpeciesT. album
PackagingBottle
Product TypeProteinase K
Quantity100 mg
Shipping ConditionDry Ice
Product LineAmbion
Unit SizeEach
Contents & Storage
Store dry powder in desiccant at –20°C.

Frequently asked questions (FAQs)

How do I isolate DNA from single bacterial colonies before PCR?

Scrape a single colony from the surface of an agar plate and transfer to 12 µL of SCL solution [10 mM Tris-HCl (pH 7.5), 1 mM EDTA, 50 µg/mL proteinase K] and incubate for 15 min at 55 degrees C. Inactivate proteinase K for 15 min at 80 degrees C and add 20 µL of deionized water. Centrifuge at 12,000 x g for 3 min. Transfer the supernatant to a new tube.

What are the recommended conditions for proteinase K treatment when isolating RNA or DNA samples?

Concentration: Generally proteinase K is used in the concentration range of 50 to 500 µg/mL at 65 degrees C in the presence of SDS (0.5-1%).

Temperature optimum: 65 degrees C; 12X more active at 65 degrees C than at 25 degrees C.

pH: Proteinase K is stable over a wide pH range (4.0 to 12.5), with optimal activity at pH 6.5 to 9.5. It is most stable at pH 8.
Inactivation: Heat inactivate Proteinase K at 80 degrees C for 15 min. Phenol extraction is the recommended method to ensure complete inactivation.

Inhibited by: PMSF (0.1 to 1.0 mM of PMSF is usually sufficient for inhibition of Proteinase K)

Not inhibited by: Proteinase K is not inactivated by metal ions, chelating agents (e.g., EDTA), sulfhydryl reagents or by trypsin or chymotrypsin inhibitors. Activity can be stimulated by addition of denaturing agents (SDS and urea).