Skip to main content Skip to navigation
Purified Mouse Anti- Human Topo IIβ
Purified Mouse Anti- Human Topo IIβ

Western blot analysis of Topo IIβ on a Jurkat lysate. Lane 1: 1:1000, lane 2: 1:2000, lane 3: 1:4000 dilution of the Topo IIβ antibody.

Purified Mouse Anti- Human Topo IIβ

Immunofluorescent staining of A549 cells.  Cells were seeded in a 96 well imaging plate (Cat. No. 353219) at ~ 10 000 cells per well.  After overnight incubation, cells were stained using the methanol fix/perm protocol (see Recommended Assay Procedure) and the anti-Topisomerase IIβ  antibody.  The second step reagent was FITC goat anti mouse Ig (Cat. No. 554001).  The image was taken on a Pathway 850 imager using a 20x objective.  This antibody also stained HeLa and U2OS cells and can be used with either fix/perm protocol (see Recommended Assay Procedure).

Western blot analysis of Topo IIβ on a Jurkat lysate. Lane 1: 1:1000, lane 2: 1:2000, lane 3: 1:4000 dilution of the Topo IIβ antibody.

Immunofluorescent staining of A549 cells.  Cells were seeded in a 96 well imaging plate (Cat. No. 353219) at ~ 10 000 cells per well.  After overnight incubation, cells were stained using the methanol fix/perm protocol (see Recommended Assay Procedure) and the anti-Topisomerase IIβ  antibody.  The second step reagent was FITC goat anti mouse Ig (Cat. No. 554001).  The image was taken on a Pathway 850 imager using a 20x objective.  This antibody also stained HeLa and U2OS cells and can be used with either fix/perm protocol (see Recommended Assay Procedure).

Product Details
Down Arrow Up Arrow


BD Transduction Laboratories™
Human (QC Testing)
Mouse IgG1
Human Topo IIβ aa. 1281-1494
Bioimaging, Western blot (Routinely Tested), Immunofluorescence (Tested During Development)
180 kDa
250 µg/ml
AB_398952
Aqueous buffered solution containing BSA, glycerol, and ≤0.09% sodium azide.
RUO


Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. Store undiluted at -20°C.

Recommended Assay Procedures

Methanol Procedure for a 96 well plate:

Remove media from wells. Add 100 µl/well fresh 3.7% Formaldehyde in PBS. Incubate for 10 minutes at room temperature (RT). Flick out and add 100 µl/well 90% methanol. Incubate for 5 minutes at RT. Flick out and wash twice with PBS. Flick out PBS and add 100 µl/well blocking buffer (3% FBS in PBS). Incubate for 30 minutes at RT. Flick out and add diluted antibody (diluted in blocking buffer). Incubate for 1 hour at RT. Wash three times with PBS. Flick out PBS and add second step reagent. Incubate for 1 hour at RT. Wash three times with PBS. Image sample.

Triton-X 100 Procedure for a 96 well plate:

Remove media from wells. Add 100 µl/well fresh 3.7% Formaldehyde in PBS. Incubate for 10 minutes at room temperature (RT). Flick out and add 100 µl/well 0.1% Triton-X 100. Incubate for 5 minutes at RT. Flick out and wash twice with PBS. Flick out PBS and add 100 µl/well blocking buffer (3% FBS in PBS). Incubate for 30 minutes at RT. Flick out and add diluted antibody (diluted in blocking buffer). Incubate for 1 hour at RT. Flick out and wash three times with PBS. Flick out and add second step reagent. Incubate for 1 hour at RT. Flick out and wash three times with PBS. Image sample.

Product Notices

  1. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  2. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
611492 Rev. 2
Antibody Details
Down Arrow Up Arrow
40/Topo IIβ

Eukaryotic DNA topoisomerase II, a ubiquitous ATP-dependent type II topoisomerase, is an essential nuclear enzyme in DNA replication and transcription, chromatin segregation, and cell cycle progression. Topoisomerases transiently break a pair of complementary strands in double-stranded DNA to form a gate for the passage of duplex DNA. Two isoforms of DNA topoisomerase II have been identified: topo IIα and topo IIß. These exhibit a high degree of homology, except for some divergence in the C-terminal region. Both contain multiple bipartite nuclear localization sequences (NLS) that mediate their subnuclear localization. Topo IIα levels rise during late S phase and peak in G2-M, whereas topo IIß levels remain constant throughout the cell cycle. In addition, topo IIα is expressed in proliferating cells, while topo IIß is expressed in a wide range of tissues. Although the exact role of these two isoforms during cell proliferation is not known, the differences in cellular expression implicate different physiological roles. Both isoforms may also be important targets for anticancer agents that exert cytotoxicity in proliferating cells via stabilization of a topo II-DNA complex.

This antibody is routinely tested by Western blot analysis and immunofluorescent imaging. Other applications were tested at BD Biosciences Pharmingen during antibody development only.

611492 Rev. 2
Format Details
Down Arrow Up Arrow
Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
611492 Rev.2
Citations & References
Down Arrow Up Arrow

Development References (5)

  1. Brown MS, Holden JA, Rahn MP, Perkins SL. Immunohistochemical staining for DNA topoisomerase IIa in Hodgkin's disease. Am J Clin Pathol. 1998; 109(1):39-44. (Biology). View Reference
  2. Herzog CE, Holmes KA, Tuschong LM, Ganapathi R, Zwelling LA. Absence of topoisomerase IIbeta in an amsacrine-resistant human leukemia cell line with mutant topoisomerase IIalpha. Cancer Res. 1998; 58(23):5298-5300. (Biology). View Reference
  3. Shain KH, Landowski TH, Dalton WS. Adhesion-mediated intracellular redistribution of c-Fas-associated death domain-like IL-1-converting enzyme-like inhibitory protein-long confers resistance to CD95-induced apoptosis in hematopoietic cancer cell lines. J Immunol. 2002; 168(5):2544-2553. (Clone-specific: Western blot). View Reference
  4. Suzuki H, Tomida A, Tsuruo T. Dephosphorylated hypoxia-inducible factor 1alpha as a mediator of p53-dependent apoptosis during hypoxia. Oncogene. 2001; 20(41):5779-5788. (Clone-specific: Western blot). View Reference
  5. Tsai-Pflugfelder M, Liu LF, Liu AA, et al. Cloning and sequencing of cDNA encoding human DNA topoisomerase II and localization of the gene to chromosome region 17q21-22. Proc Natl Acad Sci U S A. 1988; 85(19):7177-7181. (Biology). View Reference
View All (5) View Less
611492 Rev. 2

Please refer to Support Documents for Quality Certificates


Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.