产品信息

Highly characterized for more consistent performance, Proteinase K is a subtilisin-related serine protease that will hydrolyze a variety of peptide bonds. Proteinase K is active in a wide range of temperatures and buffers with optimal activity between 20 and 60°C and a pH between 7.5 and 12.0 (1, 2). Activity is stimulated when up to 2% SDS or up to 4 M urea are included in the reaction (3). Calcium is important for thermostability of Proteinase K but it is not required for catalysis, therefore Proteinase K is also active in buffers containing chelating agents such as EDTA (4).

Lot-to-Lot Variability: Competitor Comparison

Activity Measurements: Competitor Comparison

Protein Concentration
20 mg/ml, approximately, as determined by UV absorption at 280 nm.

产品来源

Engyodontium album (Tritirachium album)

产品类别:

Proteases Products,

Nucleic Acid Purification Products

• 产品组分信息

  本产品提供以下试剂或组分：

  NEB #	名称	组分货号	储存温度	数量	浓度
  P8107S     -20       Proteinase K, Molecular Biology Grade P8107SVIAL -20 2 x 1 ml 800 units/ml

• 特性和用法

  单位定义

  One unit will digest urea-denatured hemoglobin at 37°C (pH 7.5) per minute to produce equal absorbance as 1.0 μmol of L-tyrosine using Folin & Ciocalteu’s phenol reagent (6).

  贮存溶液

  20 mM Tris-HCl
  1 mM CaCl₂
  50% Glycerol
  pH 7.4 @ 25°C

  热失活

  95°C for 10 minutes

  分子量

  理论上的: 28.9 kDa

  单位活性检测条件

  0.5–2 μg of Proteinase K is incubated with 2% denatured hemoglobin solution for 10 minutes at 37°C (pH 7.5). After precipitation, neutralization and addition of Folin & Ciocalteu’s phenol reagent, absorbance of soluble cleavage products are measured at 750 nm. Absorbance is compared to a standard curve of L-tyrosine absorbance prepared similarly.

• 优势和特性

  应用特性

  • Isolation of plasmid and genomic DNA 
  • Isolation of RNA
  • Inactivation of RNases, DNases and enzymes in reactions
  • Removal of enzymes from DNA to improve cloning efficiency (5)
  • PCR purification

• 注意事项

  1. Active in a wide range of buffers, including all NEB-specific restriction endonuclease buffers. It is highly active between pH 7.5 and 12 and temperatures 20-60°C. Proteinase K is also active in chelating agents such as EDTA and activity is stimulated in up to 2% SDS or 4M urea.
  2. Proteinase K is stable for at least 2 years at –20°C. No loss of activity is observed after 10 freeze-thaw cycles.
  3. Reaction Conditions
     Proteinase K is active in a wide range of buffers including all NEB specific restriction endonuclease buffers. It is highly active between pH 7.5 and 12.0 and temperatures between 20 and 60°C (1,2). Proteinase K is also active in chelating agents such as EDTA (4) and activity is stimulated in up to 2% SDS or 4 M urea (3).

• 参考文献

  1. Bajorath, J. et al. (1988). Biochimica et Biophysica Acta. 954, 176-182.
  2. Ebeling, W. et al. (1974). Eur. J. Biochem. 47, 91-97.
  3. Hilz, H. et al. (1975). Eur. J. Biochem. 56, 103-108.
  4. Bajorath, J. et al. (1988). Eur. J. Biochem. 176, 441-447.
  5. Crowe, J.S. et al, (1991). Nucleic Acids Research. 91, 184.
  6. Anson, M.L. (1939). J. Gen. Physiol. 22, 79-89.
  7. Pace, C.N. et al. (1995). Protein Sci. 4, 2411-2423.

操作说明、说明书 & 用法

• 操作说明

  1. General protocol to release nucleic acids prior to capillary or gel electrophoresis using Proteinase K (P8107)
  2. Protocol to cleanup DNA Glucosylation/restriction digest and Proteinase using Proteinase K (P8107)
  3. Protocol to purify PCR products in preparation for cloning using Proteinase K (P8107)
  4. Using recombinant Cas9 nuclease to assess locus modification in genome editing experiments (#M0386)

FAQs & 问题解决指南

• FAQs

  1. The protocol I am using requires a µg/ml concentration of Proteinase K. What is the protein concentration of Proteinase K, Molecular Biology Grade?
  2. What buffer should I use for Proteinase K?
  3. If I need to dilute Proteinase K, Molecular Biology Grade, how should I do this?
  4. Why is the unit definition assay for Thermolabile Proteinase K (NEB #P8111) different from the unit definition assay for Proteinase K, Molecular Biology Grade (NEB # P8107)?
  5. What is the activity if Proteinase K, Molecular Biology Grade if subjected to the unit definition assay used for Thermolabile Proteinase K?