Human TOP2B Antibody

Catalog #: AF6348 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF6348
AF6348-SP

Detection of Human TOP2B by Western Blot.

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human TOP2B Antibody Summary

Specificity

Detects human TOP2B in Western blots.

Source

Polyclonal Sheep IgG

Purification

Antigen Affinity-purified

Immunogen

E. coli-derived recombinant human TOP2B
aa 1187-1621
Accession # Q02880

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

0.2 µg/mL

See below

Knockout Validated

TOP2B is specifically detected in HEK293 human embryonic kidney parental cell line but is not detectable in TOP2B knockout HEK293 cell line.

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot

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Detection of Human TOP2B by Western Blot. Western blot shows lysates of K562 human chronic myelogenous leukemia cell line. Gels were loaded with 30 µg of whole cell lysate (WCL), 20 µg of cytoplasmic (Cyto), and 10 µg of nuclear extracts (Nuc). PVDF Membrane was probed with 0.2 µg/mL of Sheep Anti-Human TOP2B Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6348) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for TOP2B at approximately 185 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group1.

Knockout Validated

Western Blot Shows Human TOP2B Antibody Specificity by Using Knockout Cell Line.

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Western Blot Shows Human TOP2B Specificity by Using Knockout Cell Line. Western blot shows lysates of HEK293 human embryonic kidney parental cell line and TOP2B knockout HEK293 cell line (KO). PVDF membrane was probed with 0.2 µg/mL of Sheep Anti-Human TOP2B Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6348) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for TOP2B at approximately 245 kDa (as indicated) in the parental HEK293 cell line, but is not detectable in knockout HEK293 cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: TOP2B

TOP2B (DNA Topoisomerase II beta ) is a 180‑185 kDa member of the type IIA subfamily, topoisomerase family of molecules. It is ubiquitously expressed, and represents the larger of two known topoisomerases (the 2A/ alpha -form being 170 kDa in size). TOP2A is essential for cell division, while TOP2B is active postmitotically. In an ATP-dependent manner, homodimeric TOP2B relieves torsional stress created in DNA during transcription, or as a consequence of replication. TOP2B first induces cleavage of double-stranded DNA, creating a space that allows for the physical repositioning of chromatin and a reduction in tension. This is followed by closure and ligation of the cleaved ends to recreate the original DNA structure. Human TOP2B is 1626 amino acids (aa) in length. It contains an ATPase domain (aa 101‑201), an NES (aa 1034‑1044), more that 30 Ser/Thr phosphorylation sites and a C-terminal DTHCT region (aa 1508‑1611). There is one splice variant that shows a deletion of aa 24‑28. Over aa 1187‑1621, human TOP2B shares 91% aa identity with mouse TOP2B.

Long Name

DNA Topoisomerase II beta

Entrez Gene IDs

7155 (Human); 21974 (Mouse)

Alternate Names

antigen MLAA-44; DNA topoisomerase 2-beta; DNA topoisomerase II beta; DNA topoisomerase II, 180 kD; DNA topoisomerase II, beta isozyme; EC 5.99.1.3; TOP2B; top2beta; TOPIIB; topo II beta; topoisomerase (DNA) II beta (180kD); topoisomerase (DNA) II beta 180kDa; topoisomerase II beta; topoisomerase IIb; U937 associated antigen