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TOP1 cell lysate

Human TOP1 Baculovirus-Insect Overexpression Lysate

Gene Names
TOP1; TOPI
Applications
Western Blot
Synonyms
TOP1; Human TOP1 Baculovirus-Insect Overexpression Lysate; Human TOP1 Insect Cell Lysate (WB positive control); TOP1 cell lysate
Ordering
For Research Use Only!
Host
Baculovirus-Insect Cells
Form/Format
1x Sample Buffer (1x modified RIPA buffer+1x SDS loading buffer).
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
Sequence Positions
594aa
Sequence
Lys191-Phe765
Applicable Applications for TOP1 cell lysate
Western Blot (WB)
Application Notes
WB: Use at an assay dependent dilution.
Species
Human
Recommend Usage
1. Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube.2. Re-dissolve the pellet using 200uL pure water and boil for 2-5 min.
3. Store the lyophilized cell lysate at 4 degree C. After re-dissolution, recommend to aliquot it into smaller quantities and store at -80 degree C.
Sequence Construction
A DNA sequence encoding the human TOP1 (NP_003277.1) (Lys191-Phe765) was expressed with a polyhistidine tag at the N-terminus.
Preparation Method
Cell lysate was prepared by homogenization in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors. Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Preparation and Storage
Store at 4 degree C for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80 degree C for up to twelve months. Avoid repeated freeze-thaw cycles.
Samples are stable for up to twelve months from date of receipt.
Related Product Information for TOP1 cell lysate
This Human TOP1 overexpression lysate was created in Baculovirus-Insect Cells and intented for use as a Western blot (WB) positive control. Purification of TOP1 protein from the overexpression lysate was verified.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
90,726 Da
NCBI Official Full Name
DNA topoisomerase 1
NCBI Official Synonym Full Names
topoisomerase (DNA) I
NCBI Official Symbol
TOP1
NCBI Official Synonym Symbols
TOPI
NCBI Protein Information
DNA topoisomerase 1; type I DNA topoisomerase
UniProt Protein Name
DNA topoisomerase 1
Protein Family
UniProt Gene Name
TOP1
UniProt Entry Name
TOP1_HUMAN

NCBI Description

This gene encodes a DNA topoisomerase, an enzyme that controls and alters the topologic states of DNA during transcription. This enzyme catalyzes the transient breaking and rejoining of a single strand of DNA which allows the strands to pass through one another, thus altering the topology of DNA. This gene is localized to chromosome 20 and has pseudogenes which reside on chromosomes 1 and 22. [provided by RefSeq, Jul 2008]

Uniprot Description

TOP1: Releases the supercoiling and torsional tension of DNA introduced during the DNA replication and transcription by transiently cleaving and rejoining one strand of the DNA duplex. Introduces a single-strand break via transesterification at a target site in duplex DNA. The scissile phosphodiester is attacked by the catalytic tyrosine of the enzyme, resulting in the formation of a DNA-(3'-phosphotyrosyl)-enzyme intermediate and the expulsion of a 5'-OH DNA strand. The free DNA strand than undergoes passage around the unbroken strand thus removing DNA supercoils. Finally, in the religation step, the DNA 5'-OH attacks the covalent intermediate to expel the active-site tyrosine and restore the DNA phosphodiester backbone. Regulates the alternative splicing of tissue factor (F3) pre-mRNA in endothelial cells. A chromosomal aberration involving TOP1 is found in a form of therapy-related myelodysplastic syndrome. Translocation t(11;20)(p15;q11) with NUP98. Belongs to the type IB topoisomerase family.

Protein type: Oncoprotein; Nucleolus; Isomerase; EC 5.99.1.2

Chromosomal Location of Human Ortholog: 20q12-q13.1

Cellular Component: nucleoplasm; nuclear chromosome; nucleolus; perikaryon; nucleus

Molecular Function: DNA topoisomerase (ATP-hydrolyzing) activity; protein binding; DNA binding; chromatin binding; DNA topoisomerase type I activity

Biological Process: circadian rhythm; response to drug; chromatin remodeling; DNA topological change; viral reproduction; programmed cell death; circadian regulation of gene expression; DNA replication; phosphorylation; embryonic cleavage; chromosome segregation

Research Articles on TOP1

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Product Notes

The TOP1 top1 (Catalog #AAA8115705) is a Cell Lysate produced from Baculovirus-Insect Cells and is intended for research purposes only. The product is available for immediate purchase. The immunogen sequence is 594aa. AAA Biotech's TOP1 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB). WB: Use at an assay dependent dilution. Researchers should empirically determine the suitability of the TOP1 top1 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. The amino acid sequence is listed below: Lys191-Phe 765. It is sometimes possible for the material contained within the vial of "TOP1, Cell Lysate" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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