Anti-Topo I-DNA Complex clone 1.1A ZooMAb^® Mouse Monoclonal

DNA topoisomerase 1 (UniProt: P11387; also known as EC:5.6.2.1, DNA topoisomerase I) is encoded by the TOP1 gene (Gene ID: 7150) in human. Topoisomerase I is a KEY^® enzyme that releases the supercoiling and torsional tension of DNA introduced during the DNA replication. It achieves this by transiently Cleaving and rejoining one strand of the DNA duplex. During the course of its normal catalytic cycle, it nicks one DNA strand, allows rotation around the intact strand, and then reseals the DNA backbone, restoring DNA integrity. The strand break occurs via formation of a transient 3′-phosphotyrosine ester bond between the active site tyrosine 723 and the DNA scissile strand backbone phosphate. This is followed by the unwinding of DNA, and by a second transesterification reaction resulting in religation of the cleaved DNA and dissociation of the topoisomerase I-DNA complex. The C-terminal domain that contains the active site tyrosine (Tyr723) lacks enzymatic activity by itself and requires its association with the 56 kDa core domain that contains the remaining catalytically relevant residues. This region also contains a number of residues that are involved in interactions with DNA. Phosphorylation of Topoisomerase I at serine 506 by casein kinase 2 is reported to increase binding to supercoiled DNA and sensitivity to camptothecin. Clone 1.1A specifically recognizes covalent topoisomerase I-DNA complexes but does not detect free Topoisomerase I or DNA. This ZooMAb^® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, Affinity^™, reproducibility, and stability over conventional monoclonals. (Ref.: Patel, AG., et al. (2016). Nucleic Acids Res. 44(6); 2816-2826; Bandyopadhyay, K., et al. (2012). PLoS One. 7(11); e50427; Punchihewa C., et al. (2007). J. Struct. Biol. 159(1); 111-121).

We are committed to bringing you greener alternative products, which adhere to one or more of The 12 Principles of Green Chemistry.This antibody is Preservative-free, produced without the harm or sacrifice of animals and exceptionally stable to allow for ambient shipping and storage if needed and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency". Click here for more information.

ZooMAb^® antibodies represent an entirely new generation of recombinant monoclonal antibodies. Each ZooMAb^® antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb^® antibodies are reliably available and ready to ship when you need them.

KLH-conjugated linear peptide corresponding to 15 amino acids surrounding phosphotyrosine 723 in human Topoisomerase I.

Quality Control Testing

Evaluated by Western Blotting in HeLa cell lysate.

Western Blotting Analysis: A 1:1,000 dilution of this antibody detected Topo I-DNA Complex in HeLa cell lysate.

Tested Applications

Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected Topo I-DNA Complex in NIH3T3 cell lysate.

Flow Cytometry Analysis: 1 µg from a representative lot detected Topo I-DNA Complex in one million fixed and permeabilized A549 cells treated with Topotecan (10 M; 20 min).

Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected Topo I-DNA Complex in A549 cells treated with Topotecan (10 M; 20 min)

Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected Topo I-DNA Complex in Human kidney tissue sections.

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.

Clone 1.1A is a mouse recombinant monoclonal antibody that specifically detects Topoisomerase I-DNA covalent complex with phosphotyrosine 723.

Purified recombinant mouse monoclonal antibody IgG, lyophilized in PBS, 5% Trehalose, normal appearance a coarse or translucent resin. The PBS/trehalose components in the ZooMAb formulation can have the appearance of a semi-solid (bead like gel) after lyophilization. This is a normal phenomenon. Please follow the recommended reconstitution procedure in the data sheet to dissolve the semi-solid, bead-like, gel-appearing material. The resulting antibody solution is completely stable and functional as proven by full functional testing. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 µL.

Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 µL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws.

Reconstitute lyophilized antibody pellet with 25μL of ultrapure water or Phosphate Buffered Saline (PBS). Please refer to our reconstitution protocol and the specific application guidance on the suggested starting dilutions and sample type.

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

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ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany

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.