Klenow Enzyme, Labeling Grade
from E. coli lysogenic NM 964, solution
For further processing only.
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Klenow Enzyme, Labeling Grade material number and pack size: Material Number Pack Size 11010492103 custom fill
- Klenow enzyme is a DNA-dependent 5'-3' polymerase with 3'-5' exonuclease activity to synthesize DNA complementary to single-stranded DNA templates. Klenow Enzyme is a DNA-dependent 5'-3' polymerase with 3'-5' exonuclease activity. It lacks the 5'-3' exonuclease activity of the native enzyme. Klenow Enzyme catalyzes the addition of mononucleotides to the 3'-OH terminus of DNA. This activity is used to synthesize DNA complementary to single-stranded DNA templates.
- Use Klenow Enzyme for:
- Random-primed DNA labeling using random oligonucleotides as primers for the incorporation of nonradioactively labeled and 32P-labeled nucleotides
- Fill-in reaction for blunt-end formation of 3'-recessed (staggered) ends
Molecular weight: 68 kD
Appearance: Colorless solution
Storage buffer: Potassium phosphate, 50 mmol/L; DTE, 1 mmol/L; glycerol, 50% (v/v); pH approximately 7.0 at +4°C
Volume activity: ≥2000 U/mL
Specific activity: ≥5000 U/mg
Unit definition: One unit is the enzyme activity which incorporates 10 nmol of total nucleotides into an acid-precipitable fraction in 30 minutes under assay conditions at +37°C with poly [d(A-T)] as primer (Richardson, C.C. & Kornberg, A. (1994) J. Biol. Chem. 244, 2996).
Nicking activity (pBR322 DNA): Not detectable in up to 50 U after 16 hours incubation at +37°C.
Purity (SDS gel electrophoresis, using up to 50 U enzyme): ≥90%
Protein: ≤0.4 mg/mL
Function test using Random Primed DNA Labeling Kit (>50% incorporation radioactive nucleotids after 30 minutes): Corresponds to specification
Stability: At -15 to -25°C within specification range for 24 months.