Proteinase K, Molecular Biology Grade |NEB酶试剂 New England Biolabs

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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 ComparisonProteinase K, Molecular Biology Grade |
Activity measurements from three NEB lots of Proteinase K were compared to three lots from another vendor. Reported activity for all NEB lots is 800 Units/ml ± 10% (indicated as a black line with error bars). Direct measurement of the competitor’s material using an assay reported on the vendor’s literature, demonstrated a dramatic over-assessment of all three lots.

Activity Measurements: Competitor ComparisonProteinase K, Molecular Biology Grade |
Activity measurements from a single NEB lot were compared to a single lot from seven other vendors. Proteinase K from NEB is sold at 800 Units/ml ± 10% (indicated as a black line with error bars), whereas most other vendors sell Proteinase K as > 800 or > 600 Units/ml. While Proteinase K from some vendors was close to the stated activity values, others varied significantly from the expected amount.


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?