Topo IIalpha (AA 1245-1361) antibody

Details for Product No. ABIN968474
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Antigen
Epitope
AA 1245-1361
(13), (3), (2), (1), (1), (1), (1), (1)
Reactivity
Human
(23), (3), (2), (1)
Host
Mouse
(23)
Clonality (Clone)
Monoclonal ()
Conjugate
Un-conjugated
(1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1)
Application
BioImaging (BI), Western Blotting (WB), Immunofluorescence (IF)
(14), (11), (10), (6), (3), (1)
Pubmed 4 references available
Quantity 50 μg
Options
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Catalog No. ABIN968474
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Immunogen Human Topo IIalpha
Clone TFR-01
Isotype IgG1
Characteristics 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 us for technical protocols.
Purification Purified from tissue culture supernatant or ascites by affinity chromatography.
Alternative Name Topo II alpha
Background 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. These enzymes 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 IIalpha and topo IIß. These have 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 IIalpha levels rise during late S phase and peak in G2-M, whereas topo IIß levels remain constant throughout the cell cycle. In addition, topo IIalpha is expressed in proliferating cells, while topo IIß is expressed in many tissues. The exact role of these two isoforms during cell proliferation is not known, however 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.
Molecular Weight 170 kDa
Application Notes 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.
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.
Comment

Related Products: ABIN968535, ABIN967389

Restrictions For Research Use only
Format Liquid
Concentration 250 µg/ml
Buffer Aqueous buffered solution containing BSA, glycerol.
Preservative Sodium azide
Precaution of Use This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Storage -20 °C
Supplier Images
anti-Topo IIalpha (AA 1245-1361) antibody Western blot analysis of Topo IIalpha on a HeLa lysate. Lane 1: 1:500, lane 2: 1:1000, lane 3: 1:2000 dilution of the Topo IIalpha antibody.
anti-Topo IIalpha (AA 1245-1361) antibody (2) Immunofluorescent staining of HeLa cells. Cells were seeded in a 96 well imaging plate at ~ 10 000 cells per well. After overnight incubation, cells were stained using the methanol fix/perm protocol and the anti-Topo IIalpha antibody. The second step reagent was FITC goat anti mouse Ig. The image was taken on a Pathway 850 imager using a 20x objective. This antibody also stained A549 and U2OS cells and can be used with either fix/perm protocol.
Product cited in: Tsai-Pflugfelder, Liu, Liu et al.: "Cloning and sequencing of cDNA encoding human DNA topoisomerase II and localization of the gene to chromosome region 17q21-22." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 85, Issue 19, pp. 7177-81, 1988 (PubMed).

Brown, Holden, Rahn et al.: "Immunohistochemical staining for DNA topoisomerase IIa in Hodgkin's disease." in: American journal of clinical pathology, Vol. 109, Issue 1, pp. 39-44, 1998 (PubMed).

Herzog, Holmes, Tuschong et al.: "Absence of topoisomerase IIbeta in an amsacrine-resistant human leukemia cell line with mutant topoisomerase IIalpha." in: Cancer research, Vol. 58, Issue 23, pp. 5298-300, 1998 (PubMed).

Mirski, Gerlach, Cole: "Sequence determinants of nuclear localization in the alpha and beta isoforms of human topoisomerase II." in: Experimental cell research, Vol. 251, Issue 2, pp. 329-39, 1999 (PubMed).

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