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Delight in the flexibility of our product offerings, now available in a glycerol-free version, meticulously crafted to seamlessly integrate with your freeze-drying systems. 

Additionally, we present a lyophilized version, a practical solution ensuring stable transportation at room temperature, thereby notably mitigating shipping costs, especially during long-distance transits.

Cat. No.: T4PK3-500 (for 500U)

Cat. No.: T4PK3-2k (for 2KU)

Cat. No.: T4PK3-10k (for 10KU)

Description

T4 Polynucleotide Kinase (3' phosphatase minus), abbreviated as T4 PNK (3' phosphatase minus), is a type of polynucleotide 5'-hydroxyl kinase. It catalyzes the transfer of the γ-phosphate group of ATP to the 5'-hydroxyl of single-stranded or double-stranded DNA, RNA, oligonucleotides, or nucleotides with a 3'-phosphate group. Other NTPs can also undergo the same reaction: 5'-OH + NTP → 5'-P + NDP.

Compared to T4 Polynucleotide Kinase, T4 Polynucleotide Kinase (3' phosphatase minus) lacks the 3' phosphatase activity, meaning it cannot catalyze the dephosphorylation of the 3' end of phosphorylated polynucleotides: 3'-P → 3'-OH + Pi (optimal pH is around 5.9).

T4 PNK (3' phosphatase minus) is particularly suitable for converting OH-DNA/RNA-p to p-DNA/RNA-p or converting Np to pNp through catalysis.

Applications

• 5' end labeling of oligonucleotides, DNA, or RNA.
• Phosphorylation of the 5' end of oligonucleotides, DNA, or RNA to facilitate subsequent ligation reactions.
• Phosphorylation of 3'-phosphorylated nucleotides to generate pNp substrates that can be ligated to the 3' end of DNA or RNA.
• 5' end labeling of 3'-phosphorylated oligonucleotides.

Source

Expressed in Escherichia coli, the expression gene originates from a mutant of T4 Polynucleotide Kinase from T4 bacteriophage.

Enzyme Activity Definition

One unit is defined as the amount of enzyme that catalyzes the incorporation of 1 nmol of phosphate from ATP to 5'-OH DNA in 30 minutes at 37°C.

Purity

Does not contain DNA endonucleases, exonucleases, or RNAse.

Enzyme Storage Solution

10 mM Tris-HCl (pH 7.4), 50 mM KCl, 0.1 mM EDTA, 1 mM DTT, 0.1 µM ATP, 50% glycerol.

T4 PNK Buffer (10X)

700 mM Tris-HCl (pH 7.6 at 25°C), 100 mM MgCl2, 50 mM DTT.

Inactivation or Inhibition

T4 Polynucleotide Kinase (3' phosphatase minus) can be inactivated by heating at 75°C for 10 minutes or 65°C for 20 minutes. Addition of EDTA can also deactivate T4 Polynucleotide Kinase (3' phosphatase minus). Metal chelators, phosphate ions, ammonium ions, KCl, and NaCl concentrations exceeding 50 mM significantly inhibit the activity of T4 Polynucleotide Kinase (3' phosphatase minus).

Precautions

• DNA obtained from ammonium salt precipitation should not be used in labeling reactions with T4 Polynucleotide Kinase (3' phosphatase minus). Ammonium salts strongly inhibit the enzymatic activity of T4 Polynucleotide Kinase (3' phosphatase minus).
• The enzyme should be stored in an icebox or on ice during use, and after use, it should be immediately stored at -20°C.
• This product is intended for scientific research purposes by professionals only and should not be used for clinical diagnosis or treatment, food or drug purposes, or stored in residential households.
• For your safety and health, please wear laboratory attire and disposable gloves when handling.

Storage

The minimum shelf life is 2 years at -20°C.

Related:

• T4 Polynucleotide Kinase Kit
• T4 Polynucleotide Kinase
• T4 Polynucleotide Kinase (3' phosphatase minus)