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AB266340

Human TOP2B (Topoisomerase II beta) knockout HEK-293T cell line

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TOP2B KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 13 bp deletion in exon 3 and 17 bp deletion in exon 3.

View Alternative Names

Antigen MLAA 44, DNA topoisomerase 2-beta, DNA topoisomerase II, DNA topoisomerase II 180 kD, DNA topoisomerase II beta, DNA topoisomerase II beta isozyme, TOP IIB, TOP2B_HUMAN, Top 2, Top2 beta, Topo II beta, Topoisomerase (DNA) II beta, Topoisomerase (DNA) II beta 180kDa, Topoisomerase IIb, U937 associated antigen, beta isozyme

3 Images
Western blot - Human TOP2B (Topoisomerase II beta) knockout HEK-293T cell line (AB266340)
  • WB

Lab

Western blot - Human TOP2B (Topoisomerase II beta) knockout HEK-293T cell line (AB266340)

Lanes 1- 2 : Merged signal (red and green). Green - ab125297 observed at 183 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

ab125297 was shown to react with Topoisomerase II beta/TOP2B in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266340 (knockout cell lysate ab257286) was used. Wild-type HEK-293T and TOP2B knockout HEK-293T cell lysates were subjected to SDS-PAGE. ab125297 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 μg/ml and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Topoisomerase II beta/TOP2B antibody (<a href='/en-us/products/primary-antibodies/topoisomerase-ii-beta-top2b-antibody-ab125297'>ab125297</a>) at 1 µg/mL

Lane 1:

Wild-type HEK-293T cell lysate at 40 µg

Lane 1:

Western blot - Human TOP2B (Topoisomerase II beta) knockout HEK-293T cell lysate (<a href='/en-us/products/cell-lysates/human-top2b-topoisomerase-ii-beta-knockout-hek-293t-cell-lysate-ab257286'>ab257286</a>)

Lane 1:

Western blot - Human TOP2B (Topoisomerase II beta) knockout HEK-293T cell pellet (<a href='/en-us/products/cell-lysates/human-top2b-topoisomerase-ii-beta-knockout-hek-293t-cell-pellet-ab279120'>ab279120</a>)

Lane 2:

TOP2B knockout HEK-293T cell lysate at 40 µg

Lane 2:

Western blot - Human TOP2B (Topoisomerase II beta) knockout HEK-293T cell line (ab266340)

Predicted band size: 183 kDa

Observed band size: 183 kDa

false

Sanger Sequencing - Human TOP2B (Topoisomerase II beta) knockout HEK-293T cell line (AB266340)
  • Sanger seq

Unknown

Sanger Sequencing - Human TOP2B (Topoisomerase II beta) knockout HEK-293T cell line (AB266340)

Allele-2 : 13 bp deletion in exon 3.

Sanger Sequencing - Human TOP2B (Topoisomerase II beta) knockout HEK-293T cell line (AB266340)
  • Sanger seq

Unknown

Sanger Sequencing - Human TOP2B (Topoisomerase II beta) knockout HEK-293T cell line (AB266340)

Allele-1 : 17 bp deletion in exon 3

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Form

Liquid

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Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 13 bp deletion in exon 3 and 17 bp deletion in exon 3

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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Product details

Recommended control: Human wild-type HEK293T cell line (ab255449). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.

We will provide viable cells that proliferate on revival.

This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.

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What's included?

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Properties and storage information

Gene name
TOP2B
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C
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Handling procedures

Initial handling guidelines

Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.

1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.

Subculture guidelines
  • All seeding densities should be based on cell counts gained by established methods.
  • A guide seeding density of 2x104 cells/cm2 is recommended.
  • Cells should be passaged when they have achieved 80-90% confluence.
Culture medium

DMEM (High Glucose) + 10% FBS

Cryopreservation medium

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

Topoisomerase II beta (TOP2B) is an essential enzyme that alters the topological states of DNA during transcription replication and chromatin remodeling. It accomplishes this by inducing transient double-strand breaks in the DNA helix to facilitate the passage of another DNA segment through this break before the enzyme reseals the break. TOP2B is also known as DNA topoisomerase (ATP-hydrolyzing) and has a molecular mass of approximately 180 kDa. The enzyme is widely expressed across various tissues but the highest levels are found in the brain and testis reflecting its vital role in these organs.
Biological function summary

Topoisomerase II beta contributes to the regulation of genetic processes such as transcription chromosome segregation and DNA repair. It is not a lone player; TOP2B often functions within a complex involving other proteins that assist in stabilizing the DNA topology changes it mediates. The enzyme's activity ensures proper chromatin dynamics particularly in neurons where it influences long-range transcription regulation essential for neural development and function.

Pathways

Topoisomerase II beta plays a significant role in DNA damage response and repair pathways interacting with intricate network proteins like RPA (Replication Protein A) and BRCA1. In addition TOP2B is involved in the transcriptional regulation pathway where it facilitates changes in chromatin structure necessary for the transcription machinery to access gene sequences. Its interactions with these proteins highlight its integration in cellular mechanisms maintaining genomic stability and regulating gene expression.

Topoisomerase II beta has associations with cancer and neurodegenerative diseases. Aberrations in TOP2B activity can lead to genomic instability contributing to oncogenesis particularly in types of leukemia and solid tumors. Elevated or dysfunctional activity of TOP2B is often observed alongside proteins like the tumor suppressor p53 an important regulator of cell cycle and apoptosis linking their combined malfunction to cancer progression. Additionally irregular TOP2B function is connected to Alzheimer's disease with evidence suggesting its involvement in neuronal death often alongside proteins like Tau further implicating its role in neurodegenerative pathologies.
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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Target data

See full target information DNA topoisomerase 2-beta
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Product promise

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