Ligase

Ligase

Ligase

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  • View Data Sheet

    Description:

    Glutamate-Cysteine Ligase, Modifier Subunit Human Recombinant

    Glutamate--cysteine ligase regulatory subunit, GCS light chain, Gamma-ECS regulatory subunit, Gamma-glutamylcysteine synthetase regulatory subunit, Glutamate--cysteine ligase modifier subunit, GCLM, GLCLR.

    Product # :

    ENZ-636

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    Description

    GCLM Human Recombinant produced in E. coli is a single polypeptide chain containing 298 amino acids (1-274) and having a molecular mass of 33.3kDa.GCLM is fused to a 24 amino acid His-tag at N-terminus & purified by proprietary chromatographic techniques.

    Source

    Escherichia Coli.

    Formulation

    The GCLM solution (1mg/ml) contains 20mM Tris-HCl buffer (pH8.0), 10% glycerol and 0.15M NaCl.

    Purity

    Greater than 90.0% as determined by SDS-PAGE.

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    • Introduction

      Glutamate-cysteine ligase (GCLM) is the first rate limiting enzyme of glutathione synthesis. The GCLM enzyme is comprised of 2 subunits, a heavy catalytic subunit and a light regulatory subunit. GCLM deficiency is associated with some forms of hemolytic anemia.

    • Synonyms

      Glutamate--cysteine ligase regulatory subunit, GCS light chain, Gamma-ECS regulatory subunit, Gamma-glutamylcysteine synthetase regulatory subunit, Glutamate--cysteine ligase modifier subunit, GCLM, GLCLR.

    • Physical Appearance

      Sterile Filtered colorless solution.

    • Stability

      Store at 4°C if entire vial will be used within 2-4 weeks. Store, frozen at -20°C for longer periods of time. For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA).Avoid multiple freeze-thaw cycles.

    • Amino Acid Sequence

      MGSSHHHHHH SSGLVPRGSH MGSHMGTDSR AAKALLARAR TLHLQTGNLL NWGRLRKKCP STHSEELHDC IQKTLNEWSS QINPDLVREF PDVLECTVSH AVEKINPDER EEMKVSAKLF IVESNSSSST RSAVDMACSV LGVAQLDSVI IASPPIEDGV NLSLEHLQPY WEELENLVQS KKIVAIGTSD LDKTQLEQLY QWAQVKPNSN QVNLASCCVM PPDLTAFAKQ FDIQLLTHND PKELLSEASF QEALQESIPD IQAHEWVPLW LLRYSVIVKS RGIIKSKGYI LQAKRRGS.

    ProSpec's products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.

    Gclm Human
  • View Data Sheet

    Name :

    T4 DNA

    Description:

    T4 DNA Ligase Recombinant

    DNA ligase 4, EC 6.5.1.1, DNA ligase IV, Polydeoxyribonucleotide synthase [ATP] 4.

    Product # :

    ENZ-286

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    Description

    T4 DNA Ligase 55.3kDa protein catalyzes the formation of a phosphodiester bond between juxtaposed 5' -phosphate and 3' -hydroxyl termini in duplex DNA or RNA. This enzyme will join blunt end and cohesive end termini as well as repair single stranded nicks in duplex DNA, RNA or DNA/RNA hybrids.

    Source

    E.Coli, cloned gene-30, bacteriophage T4.

    Formulation

    50% glycerol, 20mM Tris-HCl (pH-7.5), 50mM KCl, 1mM DTT and 0.1mM EDTA.

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    • Synonyms

      DNA ligase 4, EC 6.5.1.1, DNA ligase IV, Polydeoxyribonucleotide synthase [ATP] 4.

    • Physical Appearance

      400U/ul solution.

    • Stability

      Store T4 DNA Ligase at 4°C if entire vial will be used within 2-4 weeks.Store, frozen at -20°C for longer periods of time.For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA).Avoid multiple freeze-thaw cycles.

    • Applications

      Cloning of restriction enzyme generated DNA fragments.

      Cloning of PCR products.

      Joining of double-stranded oligonucleotide linkers or adaptors to DNA.

      Site-directed mutagenesis.

      Amplified fragment length polymorphism (AFLP).

      Ligase-mediated RNA detection.

      Nick repair in duplex DNA, RNA or DNA/RNA hybrids.

      Self-circularization of linear DNA

    • Reaction Conditions

      50mM Tris-HCl, pH7.5 at 25°C, 10mM MgCl2, 10mM DTT and 1mM ATP. Incubate at 16 °C

    • Inactivation

      T4 DNA Ligase can be inhibited by NaCl/KCl at a concentrations >than 200mM or by heating at 65 °C for 10 min or at 70 °C for 5 min.

    • Unit Definition

      1U is the amount of T4 DNA Ligase required to ligate > than 50% DNA fragments in a 20μl ligation reaction system, 6μg of λDNA-Hind III decomposition product reacts at 16°C for 30 minutes.

    ProSpec's products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.

    T4 Dna Ligase
  • View Data Sheet

    Description:

    Siah E3 Ubiquitin Protein Ligase 1 Human Recombinant

    Siah E3 ubiquitin protein ligase 1, Seven in absentia homolog 1, seven in absentia homolog 1 (Drosophila), E3 ubiquitin-protein ligase SIAH1, HUMSIAH, hSIAH1, Siah-1a, SIAH1A, EC 6.3.2.

    Product # :

    ENZ-640

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    Description

    SIAH1 Human Recombinant produced in E. coli is a single polypeptide chain containing 216 amino acids (90-282) and having a molecular mass of 24.1 kDa.SIAH1 is fused to a 23 amino acid His-tag at N-terminus & purified by proprietary chromatographic techniques.

    Source

    E.coli.

    Formulation

    The SIAH1 solution (0.25mg/1ml) contains 20mM Tris-HCl buffer (pH 8.0), 100mM NaCl, 1mM DTT and 40% glycerol.

    Purity

    Greater than 95% as determined by SDS-PAGE.

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    • Introduction

      E3 ubiquitin-protein ligase SIAH1 (SIAH1) belongs to the SIAH (seven in absentia homolog) family. SIAH1 is a tumor suppressor protein, which is expressed in intestinal epithelium and activated during apoptosis. SIAH1 is involved in ubiquitination and proteasome-mediated degradation of specific proteins. SIAH1 is comprised of an N-terminal RING-finger domain (required for proteolysis) and a cystein-rich C-terminal domain (which regulates oligomerization and SIAH binding to target proteins). SIAH1 causes indirect degradation of beta-catenin by way of creation of a complex with Siah-interacting protein (SIP), Skp1 and Ebi.

    • Synonyms

      Siah E3 ubiquitin protein ligase 1, Seven in absentia homolog 1, seven in absentia homolog 1 (Drosophila), E3 ubiquitin-protein ligase SIAH1, HUMSIAH, hSIAH1, Siah-1a, SIAH1A, EC 6.3.2.

    • Physical Appearance

      Sterile Filtered colorless solution.

    • Stability

      Store at 4°C if entire vial will be used within 2-4 weeks. Store, frozen at -20°C for longer periods of time. For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA).Avoid multiple freeze-thaw cycles.

    • Amino Acid Sequence

      MGSSHHHHHH SSGLVPRGSH MGSVANSVLF PCKYASSGCE ITLPHTEKAD HEELCEFRPY SCPCPGASCK WQGSLDAVMP HLMHQHKSIT TLQGEDIVFL ATDINLPGAV DWVMMQSCFG FHFMLVLEKQ EKYDGHQQFF AIVQLIGTRK QAENFAYRLE LNGHRRRLTW EATPRSIHEG IATAIMNSDC LVFDTSIAQL FAENGNLGIN VTISMC.

    ProSpec's products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.

    Siah1 Human
  • View Data Sheet

    Description:

    HECT and RLD Domain Containing E3 Ubiquitin Protein Ligase 5 Human Recombinant

    HERC5, HECT and RLD Domain Containing E3 Ubiquitin Protein Ligase 5, CEB1, Hect Domain and RLD 5, Cyclin-E-Binding Protein 1, CEBP1, HECT Domain and RCC1-Like Domain-Containing Protein 5, E3 ISG15--Protein Ligase HERC5, Probable E3 Ubiquitin-Protein Ligase HERC5, EC 6.3.2.- , EC 6.3.2.

    Product # :

    ENZ-797

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    Description

    HERC5 Human Recombinant produced in E.Coli is a single, non-glycosylated polypeptide chain containing 367 amino acids (681-1024 a.a.) and having a molecular mass of 43kDa. HERC5 is fused to a 23 amino acid His-tag at N-terminus.

    Source

    Escherichia Coli.

    Formulation

    HERC5 protein solution (1mg/ml) contains 20mM Tris-HCl buffer (pH 8.0), 0.4M Urea and 10% glycerol.

    Purity

    Greater than 90.0% as determined by SDS-PAGE.

    More Info

    • Introduction

      HECT and RLD Domain Containing E3 Ubiquitin Protein Ligase 5 (HERC5) is a member of the HERC family of ubiquitin ligases, found in a cluster of HERC family genes on chromosome 4. HERC5 is a protein with a HECT domain and 5 RCC1 repeats. The HERC5 protein localizes to the cytoplasm and perinuclear region and serves as an INF-induced E3 protein ligase that mediates ISGylation of protein targets. HERC5 exhibits antiviral activity towards HIV-1, influenza A virus and human papillomavirus. HERC5 is a major E3 ligase for ISG15 conjugation. HERC5 also serves as a positive regulator of innate antiviral response in cells induced by INF. Pro-inflammatory cytokines upregulate HERC5 in endothelial cells. HERC5 is physically connected with polyribosomes, broadly modifies recently synthesized proteins in a cotranslational fashion.

    • Synonyms

      HERC5, HECT and RLD Domain Containing E3 Ubiquitin Protein Ligase 5, CEB1, Hect Domain and RLD 5, Cyclin-E-Binding Protein 1, CEBP1, HECT Domain and RCC1-Like Domain-Containing Protein 5, E3 ISG15--Protein Ligase HERC5, Probable E3 Ubiquitin-Protein Ligase HERC5, EC 6.3.2.- , EC 6.3.2.

    • Physical Appearance

      Sterile Filtered colorless solution.

    • Stability

      Store at 4°C if entire vial will be used within 2-4 weeks. Store, frozen at -20°C for longer periods of time. For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA).Avoid multiple freeze-thaw cycles.

    • Amino Acid Sequence

      MGSSHHHHHH SSGLVPRGSH MGSFDLTVRR NHLIEDVLNQ LSQFENEDLR KELWVSFSGE IGYDLGGVKK EFFYCLFAEM IQPEYGMFMY PEGASCMWFP VKPKFEKKRY FFFGVLCGLS LFNCNVANLP FPLALFKKLL DQMPSLEDLK ELSPDLGKNL QTLLDDEGDN FEEVFYIHFN VHWDRNDTNL IPNGSSITVN QTNKRDYVSK YINYIFNDSV KAVYEEFRRG FYKMCDEDII KLFHPEELKD VIVGNTDYDW KTFEKNARYE PGYNSSHPTI VMFWKAFHKL TLEEKKKFLV FLTGTDRLQM KDLNNMKITF CCPESWNERD PIRALTCFSV LFLPKYSTME TVEEALQEAI NNNRGFG.

    ProSpec's products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.

    Herc5 Human

About Ligase:

A ligase is an enzyme that can form a new chemical bond to catalyse the fusing of two large molecules through an accompanying hydrolysis of a small pendant chemical group. Ligases use ATP to form the bonds, and is used in recombinant DNA cloning, joining two complementary fragments of nucleic acid.

Ligase Function
DNA ligase enzymes carry the EC number 6.5.1.1 (CAS number 9015-85-4) and are used for the joining of two DNA strands by repairing single-strand breaks to catalyse phosphodiester bonds. The relationship occurs between the 5' phosphate end of the donor nucleotide and the 3’ hydroxyl ends of the acceptor nucleotide.
The four-step process, which requires AMP, involves: the reorganization of activity in DNA segments or Okazaki fragments, adenyltation of a lysine residue to the active centre of the enzyme to release the pyrophosphate, transfer of AMP to the donor nucleotide’s 5’ phosphate end to form a pyrophosphate end, formation of the phosphodiester bond between the donor nucleotide and the 3' hydroxyl of the acceptor.

Ligase Interactions
Encoded by the lig gene, The E.coli DNA ligase gains energy by cleaving the nicotinamide adenine dinucleotide (NAD) to generate a phosphodiester bond, but cannot join RNA to DNA. DNA polymerase concentrates can be used to heighten the activity of the E.coli DNA ligase but the concentrates must be smaller than the fragments to be ligated.
The Escherichia virus T4 is primarily used in lab research, and can be ligated to either the cohesive or blunt ends of oligonucleotides in addition to both RNA and RNA-DNA hybrids. However, it doesn't join single-stranded nucleic acid or use NAD. T4 relies on an ATP cofactor and has an optimal temperature of 16 °C.

Ligase Mechanism
The thermostable enzyme is derived from the thermophilic bacterium and activates at far greater temperatures than other DNA ligases, underlined by half-life rates of over 1 hour at 95 °C and 48 hours at 65 °C. It permits high hybridization stringency and ligation specificity, showing activeness for 500 thermal cycles.