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Antibody System Recombinant Bovine Terminal Transferase (TdT)

Antibody System Recombinant Bovine Terminal Transferase (TdT)

Overview
This recombinant Terminal Transferase (TdT) is a template-independent DNA polymerase cloned from bovine and expressed in an optimized E. coli system. Terminal deoxynucleotidyltransferase functions as a unique and indispensable tool in molecular biology, nucleic acid engineering, and synthetic biology due to its rare ability to synthesize DNA without a complementary template strand. The enzyme catalyzes the repetitive addition of deoxynucleotides (dNTPs) directly to the 3-prime hydroxyl (3'-OH) terminus of single-stranded or double-stranded DNA molecules.
To initiate synthesis, TdT requires an oligodeoxynucleotide initiator of at least three bases acting as a primer. Incorporating specific divalent cations significantly modulates the enzyme's relative nucleotide incorporation efficiency. While magnesium (Mg2+) supports standard extension, the addition of cobalt (Co2+) into the reaction matrix maximizes the efficiency of the tailing reaction, particularly when modifying blunt or recessed 3-prime ends. This enzyme provides an essential analytical platform for 3-prime end labeling of DNA with radioactive or non-radioactive tags, homopolymer tailing of vectors for cloning, synthetic gene assembly, and terminal tracking within TUNEL assays to detect apoptosis.
 
DATASHEET

  • PRODUCT INFO

    Key Features and Performance Metrics

    • Template-Independent Extension: Catalyzes the smooth synthesis of homopolymer or heteropolymer tails on single- or double-stranded DNA fragments completely free of template direction.

    • Refined Proteomic Purity: Exhibits a structural purity exceeding 90 percent, quantified and verified via Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE) to ensure a clean baseline absent of interfering exonuclease or endonuclease contaminants.

    • Precise Molecular Mass: Features a predicted recombinant molecular weight of 45 kDa, delivering high-yield operational kinetics and excellent solution-phase structural stability.

    • Concentration and Yield Optimization: Formulated at a clear stock concentration of 0.9 mg/ml to support direct integration into routine homopolymer tailing layouts, vector construction setups, and diagnostic labeling assays.

    Technical Specifications

    • Catalog No.: YBC05501

    • Enzyme Family: Template-Independent DNA Polymerase

    • Source Species: Bovine

    • Expression System: E. coli

    • Nature: Recombinant

    • Stock Concentration: 0.9 mg/ml

    • Predicted Molecular Weight: 45 kDa

    • Validated Application: Catalytic addition of deoxynucleotides to the 3-prime hydroxyl terminus of DNA molecules

    • Endotoxin Level: Batch-specific; please contact technical services for current certificate of analysis metrics.

    Alternative Names: DNA nucleotidylexotransferase, Terminal deoxynucleotidyltransferase, Terminal addition enzyme, TDT, DNTT, EC 2.7.7.31.

    Applications and Workflow Summary

    This recombinant bovine transferase is highly optimized for performance in 3-prime end homopolymer tailing, directed labeling protocols, and molecular cloning workflows. Investigators should observe the following guidelines:

    • Primer Requirements: Ensure your reaction system includes an oligodeoxynucleotide primer substrate possessing a length of at least three bases with an available, unblocked 3-prime hydroxyl group.

    • Tailing Optimization: For maximum tailing efficiency and uniform homopolymer extension (especially when working with blunt or 3-prime recessed DNA architectures), the introduction of cobalt ions (Co2+) into the reaction mixture is highly recommended.

    • Workflow Adjustments: Optimal operational enzyme volumes, dNTP concentrations, incubation temperatures, and durations vary depending on the desired length of the nucleotide tail and should be determined empirically by the investigator.

    • Handling Precautions: To prevent physical shear stress, misfolding, or a sudden loss of polymerase activity, handle stock aliquots with care, work on ice during reaction assembly, and centrifuge the vial briefly prior to opening.

    Handling, Stability and Storage

    • Liquid Formulation: Supplied in a highly stable liquid format dissolved in 100mM KAc (pH 6.8), 2mM 2-mercaptoethanol, 0.01 percent Triton X-100, and 50 percent glycerol cryoprotectant to safeguard structural active-site dynamics.

    • Shipping Configurations: Distributed under secure, temperature-regulated parameters utilizing dry ice or chilled blue ice packs unless customized alternative shipping arrangements are established.

    • Storage Guidance:

      • Short-Term Storage: Maintain at 2 to 8 degrees C for immediate, short-term application needs (up to 1 week).

      • Long-Term Storage: For extended preservation, store continuously at -20 to -80 degrees C for a stable framework spanning 12 months from the initial date of batch receipt.

    • Operational Shelf Life: To avoid physical stress, structural denaturation, or a sudden loss of catalytic activity, utilize a manual defrost freezer and strictly avoid repeated freeze-thaw cycles.

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