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Proteinase K (Lyophilizate, Ultra Pure)

C12100
CAT NO PRODUCT NAME SIZE PRICE
C12100 Proteinase K (Lyophilizate, Ultra Pure) 1 g $312.00
C12101 Proteinase K (Lyophilizate, Ultra Pure) 10 g $2,602.00
C12102
Proteinase K (Lyophilizate, Ultra Pure)
100g $23,410.00
PDB-1000 Buffer PDB 1000ml $300.00

Introduction

Proteinase K is a stable serine protease with broad substrate specificity. It degrades many proteins in the native state even in the presence of detergents. It is not activated by metalions, chelating agents (for example, EDTA), sulfhydryl reagents, or by tripsinor chymotrypsin inhibitors. It is stable over a wide pH range (4-12.5), with optimal activity at pH 6.5–9.5. Activity can be stimulated by addition of denaturing agents (SDS and Urea). Rapid denaturation of the enzyme occurs at temperature above 70°C. Autolysis of the enzyme occurs increasingly at alkaline pH. However, Proteinase K is not completely inactivated by autolysis. Some enzyme fragments continue to maintain their complete proteolytic activity, even after extensive autolysis.

Proteinase K is frequently used in molecular biology applications to digest unwanted proteins, such as nucleases in DNA or RNA preparations from microorganisms, cultured cells, and plants. The enzyme is typically used at 50–200 μg/ml in nucleic acid preparations at pH 7.5-8.0 and 37-55°C. Incubation times vary from 30 minutes to 18 hours.

Details


Specifications

Features Specifications
Molecular Weight 29.3kDa
Isoelectric Point 8.9
Appearance White lyophilized powder
Purity 95% (SDS-PAGE analysis)
Specific activity ≥34 Units/mg protein
Temperature characteristic The effective activity temperature is 37-70°C, and the enzyme activity at 65°C is twice that at 25°C.
PH characteristics
4.0-12.0, the optimum range is ph7.5-11.5
Preservation Conditions

It is recommended to store at -20°C to ensure the stability of activity to the greatest extent (normal temperature transportation or storage will not reduce enzyme activity).

The shelf life is up to 3 years at -20°C and 2years at 2~8°C.

Usage method

Prepare 20mg/ml with the solution, add proteinase K to the digestive solution or lysate until the final concentration is 50-200ug/ml, and incubate at 55~70°C.

Proteinase K after reaction can be removed or inactivated by magnetic bead method, column method or phenol chloroform extraction. Protease K can be inactivated by incubation at 95°C for 3 minutes or 70°C for 15 minutes.

Nucleic acid residue detection

Qubit did not detect

Human DNA contamination not detected (real time PCR)

Bacterial DNA contamination not detected (16S universal primer PCR, 30 cycles)

Fungal DNA contamination not detected (ITS Primer PCR)

Nuclease detection

DNase not detected

RNase not detected

Nickase not detected

Recommended application
Nucleic acid extraction, circulating DNA extraction, virus nucleic acid extraction


Advantages
  • Not including RNase, DNase and Nickase
  • No nucleic acid residue (not detected by Qubit after extracting the whole 100mg protein K)
  • No human-source DNA contamination (not detected by Q-PCR)
  • No bacterial DNA contamination (not detected by 16S universal primer)
  • No fungal DNA contamination (not detected by 1TS universal primer)
  • Can be transported and valid for two years at normal temperature in dry condition.
  • Can be used for highly sensitive free DNA extraction, virus nucleic acid extraction, whole blood DNA extraction, etc.


Ordering information

Contents C12100
C12101 C12102
PDB-1000
Proteinase K, Lyophilizate, >30 units/mg of protein 1 g
10 g 100 g

Buffer PDB (20mM Tris, pH 7.5, 10mM CaCl2, 50% glycerol, 0.1% Preservatives)



1000 ml

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