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MABE1905-100UG
Sigma-AldrichAnti-DNA ligase I Antibody, clone 5H5
Anti-DNA ligase I, clone 5H5, Cat. No. MABE1905, is a mouse monoclonal antibody that detects DNA ligase 1 and is tested for use in Immunocytochemistry, Immunohistochemistry, and Western Blotting.
More>>Anti-DNA ligase I, clone 5H5, Cat. No. MABE1905, is a mouse monoclonal antibody that detects DNA ligase 1 and is tested for use in Immunocytochemistry, Immunohistochemistry, and Western Blotting. Less<<
DNA ligase 1 (UniProt: P18858; also known as EC:6.5.1.1, DNA ligase I, Polydeoxyribonucleotide synthase [ATP] 1) is encoded by the LIG1 gene (Gene ID: 3978) in human. DNA ligase is the enzyme that seals nicks in double-stranded DNA during DNA replication, DNA recombination and DNA repair. DNA ligase I joins adjacent Okazaki fragments generated during the lagging strand synthesis and is recruited to replication factories during the S-phase of cell cycle. The recruitment is directed replication factory-targeting sequence (RFTS), which is also shown to mediate its interaction with the proliferating cell nuclear antigen (PCNA). Its C-terminal domain contains the catalytic region and consists of a DNA binding domain (aa 262-534), an adenylation domain (aa 535-747), and an OB-fold domain. The adenylation domain contains five conserved motifs (I, III, IIIa, IV, and V) that are reported to contribute to the surface of nucleotide binding pocket. The N-terminal region is non-catalytic and participates in nuclear localization and DNA replication. Human DNA ligase I can be phosphorylated on serine 66 by casein kinase 2 in a cell cycle-dependent manner. Several other phosphorylation sites have also been reported and the enzyme exits in a hyperphosphorylated state during the M-phase. (Ref.: Vitolo, B., et al. (2005). Eur. J. Histochem. 49(4); 349-354; Ferrari, G., et al. (2003). J. Biol. Chem. 278(39); 37761-3777; Howes, TR., and Tomkinson, AE. (2012). Subcell. Biochem. 62:327-41).
References
Product Information
Format
Purified
Presentation
Purified mouse monoclonal antibody IgG1 in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.
Applications
Application
Anti-DNA ligase I, clone 5H5, Cat. No. MABE1905, is a mouse monoclonal antibody that detects DNA ligase 1 and is tested for use in Immunocytochemistry, Immunohistochemistry, and Western Blotting.
Key Applications
Immunocytochemistry
Immunohistochemistry
Western Blotting
Application Notes
Immunohistochemistry Analysis: A representative lot detected DNA ligase I in Immunohistochemistry applications (Vitolo, B., et. al. (2005). Eur J Histochem. 49(4): 349-54).
Western Blotting Analysis: A representative lot detected DNA ligase I in Western Blotting applications (Vitolo, B., et. al. (2005). Eur J Histochem. 49(4): 349-54).
Immunocytochemistry Analysis: A representative lot detected DNA ligase I in Immunocytochemistry applications (Vitolo, B., et. al. (2005). Eur J Histochem. 49(4): 349-54).
Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user
Biological Information
Immunogen
His-tagged purified human recombinant DNA ligase I overexpressed in a baculovirus/insect cell system.
Clone
5H5
Concentration
1 mg/mL. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.
Host
Mouse
Specificity
Clone 5H5 is a mouse monoclonal antibody that detects human DNA ligase I.
Isotype
IgG1κ
Species Reactivity
Human
Mouse
Species Reactivity Note
Human, Mouse. Predicted to react with Rat, Monkey based on 100% sequence homology.
~100 kDa observed; 101.74 kDa calculated. Uncharacterized bands may be observed in some lysate(s).
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Quality Assurance
Evaluated by Western Blotting in HeLa cell lysates.
Western Blotting Analysis: 1 µg/mL of this antibody detected DNA ligase I in HeLa cell lysates.
Usage Statement
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
A new monoclonal antibody against DNA ligase I is a suitable marker of cell proliferation in cultured cell and tissue section samples B Vitolo 1 , M R Lidonnici, C Montecucco, A Montecucco Eur J Histochem
49(4):
349-54
2004
The extensive characterization of the replicative human DNA ligase I (LigI) undertaken in the last decade demonstrated that the level of this protein strongly correlates with the rate of cell proliferation. This may allow to expand the repertoire of clinical biomarkers for the analysis of cell proliferation. We have produced a new monoclonal antibody (5H5) against LigI and exploited it as cell proliferation marker in Western blotting and immunofluorescence as well as in immunohistochemistry on paraffin tissue sections. The Western blot analysis showed that the LigI level detected by 5H5 antibody is high in all proliferating cells. On the contrary the protein is down regulated in resting human fibroblast and peripheral blood lymphocytes. Immunofluorescence analysis on cultured HeLa cells showed that 5H5 antibody labels all proliferating cells and displays the same staining pattern of BrdU in S-phase nuclei. Finally the analysis of serial sections of inflamed tonsils and NHL lymph nodes (either frozen or paraffin embedded) demonstrated that 5H5 marks the same population of cells as the Ki-67 antibody. Our results demonstrate that 5H5 antibody is a valuable tool for labeling proliferating cells that can be conveniently used in Western blotting, immunocytochemistry and immunohistochemistry.
