MA5-24310

antibody from Invitrogen Antibodies

Targeting: TOP2B

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-24310

Provider

Invitrogen Antibodies

Product name

TOP2B Monoclonal Antibody (681417)

Antibody type

Monoclonal

Antigen

Recombinant full-length protein

Description

In direct ELISAs and Western blots, approximately 75% cross-reactivity with recombinant mouse TOP2B and no cross-reactivity with recombinant human TOP2A is observed.

Antibody clone number

681417

Concentration

0.5 mg/mL

References

Submitted references

SPOP is essential for DNA-protein cross-link repair in prostate cancer cells: SPOP-dependent removal of topoisomerase 2A from the topoisomerase 2A-DNA cleavage complex.

Watanabe R, Maekawa M, Hieda M, Taguchi T, Miura N, Kikugawa T, Saika T, Higashiyama S
Molecular biology of the cell 2020 Mar 15;31(6):478-490

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Mutations in topoisomerase IIβ result in a B cell immunodeficiency.

Broderick L, Yost S, Li D, McGeough MD, Booshehri LM, Guaderrama M, Brydges SD, Kucharova K, Patel NC, Harr M, Hakonarson H, Zackai E, Cowell IG, Austin CA, Hügle B, Gebauer C, Zhang J, Xu X, Wang J, Croker BA, Frazer KA, Putnam CD, Hoffman HM
Nature communications 2019 Aug 13;10(1):3644

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis from lysates of HeLa human cervical epithelial carcinoma cell line, Jurkat human acute T cell leukemia cell line, MOLT-4 human acute lymphoblastic leukemia cell line, K562 human chronic myelogenous leukemia cell line, and CH-1 mouse B cell lymphoma cell line. PVDF Membrane was probed with 1 µg/mL of mouse Anti-human/mouse TOP2B Monoclonal Antibody (Product # MA5-24310) followed by HRP-conjugated Anti-mouse IgG Secondary Antibody. A specific band was detected for TOP2B at approximately 185 kDa (as indicated). This experiment was conducted under reducing conditions.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemical analysis of TOP2B was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using mouse Anti-human/mouse TOP2B Monoclonal Antibody (Product # MA5-24310) at 25 µg/mL for 3 hours at room temperature. Cells were stained using the 557-conjugated Anti-mouse IgG Secondary Antibody (red, upper pane and counterstained with DAPI (blue, lower panel). Specific staining was localized to nuclei.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Fig. 6 Heterozygous mutation in Top2b negatively affects B-cell development beyond haploinsufficiency. Expression of mutant Top2b ( n = 7) results in reduced CD19+ B cells ( a ) in peripheral blood compared to wild-type ( n = 6) or hemizygous mice ( n = 6), while T cells ( b ) are unaffected.(shown as mean +- SEM, * p < 0.05, ** p < 0.005, two-tailed Student's t test). c qPCR analysis of kappa-deleting recombination excision circles demonstrates that B cells isolated from Top2b +/EE587E ( n = 7) or Top2b +/- mice ( n = 7) have a less robust replication history compared to wild-type mice ( n = 8 mice), shown as mean +- SEM, while T-cell proliferation as measured by T-cell receptor excision circle (TREC) formation is unaffected ( d ), ( n = 6 WT, 6 mutant and 4 hemizygous mice, performed with technical duplicates shown as mean +- SEM). * p < 0.05, ** p < 0.005, one-way ANOVA. e - g Comet assay shows increased DNA strand breaks in B cells ( e , f ) isolated from Top2b +/EE587E mice compared to wildtype, expressed as comet tail length. T cells ( g ) are not affected ( n = 3 mice per group, 10 images per mouse (X40); * p < 0.05; **** p < 0.0001, by Kruskal-Wallis test ( f ), or nonsignificant (n.s.) by Student's t test ( g )). h , i Expression of Top2b is elevated in wild-type B cells compared to wild-type T cells by RT-qPCR ( h , n = 5 mice, performed in technical triplicates, shown as mean +- SEM, * p < 0.05, two-tailed Student's t test), and immunoblot ( i , protein isolated fr