T4427

Terminal Transferase from calf thymus

buffered aqueous glycerol solution

• Sinónimos: TdT, Terminal Deoxynucleotidyl Transferase
• Número CAS: 9027-67-2
• UNSPSC Code: 12352204
• EC Number: 232-819-6
• MDL number: MFCD00166324
• Número CE: 2.7.7.31 (BRENDA, IUBMB)

Propiedades

• grade: Molecular Biology
• form: buffered aqueous glycerol solution
• mol wt: 60 kDa
• concentration: >5000 U/mL
• UniProt accession no.: P06526
• foreign activity: Exonuclease and endonuclease, free
• storage temp.: −20°C
• Gene Information: cow ... DNTT(281120)

Descripción

General description

Bovine terminal transferase (TdT) is a primer-dependent polymerase that catalyzes the addition of deoxynucleotides to the 3′-OH terminus of DNA molecules with the release of inorganic phosphate. TdT reacts preferentially with either single-stranded DNA molecules or double-stranded-DNA with 3′ overhangs, but procedures have been developed to label blunt ends or 3′-recessive ends. In a reaction mixture, the divalent ion (Co²⁺, Mn²⁺, Mg²⁺) will influence purine and pyrimidine polymerization rate. Activities of TdT are also affected by the bases (dATP, dCTP, dGTP and dTTP) present.

Application

Suitable for:

• Addition of homopolymers to vectors, inserts and cDNA for cloning
• Labeling the 3′-end of double- and single-stranded DNA with non-radioactive or radioactive labels
• Carrying out in vitro mutagenesis by adding single nucleotides to DNA
• Use in TUNEL assays

Analysis Note

Activity assay uses 200 mM potassium cacodylate, pH 7.2, 4 mM MgCl₂, 1 mM 2-mercaptoethanol, 1 mM ³H-dATP, 70 μM d(pT)₆, 37 °C.

Other Notes

One unit will incorporate 1 nanomole of dATP into acid-precipitable material in one hour at 37 °C using d(pT)₆ as primer.

Terminal transferase is supplied as a solution of 50 mM potassium phosphate (pH 7.4), 1 mM 2-mercaptoethanol and 50% glycerol (v/v).

Información de seguridad

• pictograms: GHS08
• signalword: Danger
• hcodes: H334
• pcodes: P261 - P284 - P501
• Hazard Classifications: Resp. Sens. 1
• Clase de almacenamiento: 11 - Combustible Solids
• wgk: WGK 3
• flash_point_f: Not applicable
• flash_point_c: Not applicable
• ppe: Eyeshields, Gloves, multi-purpose combination respirator cartridge (US)