DNA topoisomerase 1, EC 188.8.131.52, DNA topoisomerase I, TOP1, Scl-70.
TOP1 is an important nuclear enzyme that interconverts supercoiled DNA to the necessary topological conformations for standard DNA replication and transcription. TOP1 is the target antigen for TOP1 autoantibodies. TOP1 antibodies are a specific marker in scleroderma patients (specificity 98-100%) and are related with the existence of diffuse skin involvement and pulmonary fibrosis. In human tissues top1 enzyme is primarily synthesized as a protein with a molecular weight of 100-kDa. Most of this precursor is then proteolytically processed to a 70-kDa size, from which the TOP1 antigen has derived its name.
Recombinant TOP1 70kDa protein is an enzyme fragment having a molecular mass of 69,199 Dalton (pH 9.4). TOP1 70kDa protein is fused to a hexa-histidine purification tag.
Sf9 insect cells.
TOP1 70kDa is supplied in 16mM HEPES buffer pH-8.0, 400mM NaCl, and 20% glycerol.
1. Binds IgG-type human auto-antibodies.
2. Standard ELISA test (checkerboard analysis of positive/negative sera panels, including CDC international reference sera).
Western blot with Scleroderma patient sera or monoclonal anti-hexa-His-tag antibody.
0.5-0.8 µg/ml (depending on the type of ELISA plate and coating buffer). Suitable for biotinylation and iodination.
Store at 4°C if entire vial will be used within 2-4 weeks. Store, frozen at -20°C for longer periods of time.
Avoid multiple freeze-thaw cycles.
Greater than 80% as determined by SDS-PAGE.
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.