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Top2a

Manufactured by Santa Cruz Biotechnology
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TOP2A is a laboratory equipment product that functions as a topoisomerase II alpha enzyme. This enzyme is responsible for regulating the topology of DNA during cellular processes such as transcription, replication, and chromosome segregation. The core function of TOP2A is to introduce temporary double-strand breaks in DNA and then reseal them, allowing the DNA to unwind and relax.

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Lab products found in correlation

Top2b, by Santa Cruz Biotechnology (3 mentions) Secondary antibodies with, by LI COR (2 mentions) Image studio version 5, by LI COR (2 mentions) Odyssey clx imaging system, by LI COR (2 mentions) Conjugated 680nm and 800nm fluorophores, by LI COR (2 mentions) Kdm4b jmjd2b, by Fortis Life Sciences (2 mentions) Mdr1 abcb1, by Cell Signaling Technology (2 mentions) Vinculin, by Merck Group (1 mentions) Anti mouse hrp, by GE Healthcare (1 mentions) Protein g dynabead, by Thermo Fisher Scientific (1 mentions) Anti his, by Merck Group (1 mentions) Cenpf, by Abcam (1 mentions) Hrp protein a, by BD (1 mentions) Cdc20, by Santa Cruz Biotechnology (1 mentions) Flag antibody, by Merck Group (1 mentions) Protease inhibitor cocktail, by Merck Group (1 mentions) Anti flag m2, by Merck Group (1 mentions) Anti ha 11, by Fortrea (1 mentions) β actin, by Santa Cruz Biotechnology (1 mentions) Nolc1, by Proteintech (1 mentions)

5 protocols using top2a

1

Western Blot Profiling of Nuclear Factors

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Nuclear or Whole Cell Extracts were prepared, and protein concentrations were measured. 15ug of protein was run on a Bis-Tris gel, transferred at 200mA for 2 hours and blocked in 5% non-fat milk for 1 hour. Blots were incubated with primary antibodies overnight and then washed 3x in TBST. Blots were then incubated with secondary antibodies with conjugated −680nm and −800nm fluorophores (Licor, 1:25,000) for 45 minutes at room temperature, washed 3x, and then developed on an Odyssey® CLx Imaging System (Licor, Lincoln, NE United States). KDM4B/JMJD2B (Bethyl Laboratories A301–478A, 1:1000), TOP2A (Santa Cruz Biotech sc-3659, clone F-12, 1:1000), TOP2B (Santa Cruz Biotech H-286, sc-13059, 1:1000), MDR1/ABCB1 (Cell Signalling Technologies, 12683S, 1:1000), RNF2 (Cell Signalling Technologies, 5694S, 1:1000), H3 (Epicypher, SKU: 13–0001, 1:5000) We were unable to validate any of the available KAT6B antibodies. Licor software (Image Studio version 5.2.5) was used for the following: Image adjustment was limited to exposure (brightness and contrast; all lanes done equally and simultaneously) followed by cropping and exporting in .png format. No additional image manipulation was performed.
Seoane J.A., Kirkland J.G., Caswell-Jin J.L., Crabtree G.R, & Curtis C. (2019). Chromatin Regulators Mediate Anthracycline Sensitivity in Breast Cancer. Nature medicine, 25(11), 1721-1727.
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2

Western Blot and Immunoprecipitation Assay

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Whole protein extracts were obtained by lysing cells in TNN buffer (50mM Tris (pH 7.5), 120mM NaCl, 5mM EDTA, 0.5% NP40, 10mM Na4P2O7, 2mM Na3VO4, 100mM NaF, 1mM PMSF, 1mM DTT, 10mM β-glycerophosphate, protease inhibitor cocktail (Sigma)). Protein lysates or purified protein were separated by SDS-PAGE, transferred to a PVDF-membrane and detected by immunoblotting with the first and secondary antibodies: β-actin (Santa Cruz Biotechnology, sc-47778) 1:5000, B-MYB (clone LX015.1, [42 (link)] 1:5, anti-HA.11 (Covance, MMA-101P) 1:1000, anti-FLAG M2 (Sigma, F3165) 1:5000, anti-His (Sigma, H1029) 1:2000, Vinculin (Sigma, V9131) 1:10000, TOP2A (Santa Cruz Biotechnology, sc-365916) 1:1000, CDC20 (Santa Cruz Biotechnology, sc-5296) 1:500, YAP (Santa Cruz Biotechnology, sc-10199) 1:1000, p-YAP(S127A) (Cell Signaling Technology, 4911) 1:1000, LIN9 (Bethyl, A300-BL2981), NUSAP1 (Geert Carmeliet) 1:1000, CENPF (Abcam, ab-5) 1:1000, anti-mouse-HRP (GE healthcare, NXA931) 1:5000 and HRP Protein A (BD Biosciences, 610438) 1:5000. For immunoprecipitation of FLAG-tagged proteins, protein G dynabeads (Thermo Fisher Scientific) were first coupled with 1 μg FLAG-antibody (Sigma, F3165) and then immunoprecipitated with 1mg whole cell lysate. After five times of washing with TNN, proteins were separated by SDS-PAGE and detected by immunoblotting using the desired antibodies.
Gründl M., Walz S., Hauf L., Schwab M., Werner K.M., Spahr S., Schulte C., Maric H.M., Ade C.P, & Gaubatz S. (2020). Interaction of YAP with the Myb-MuvB (MMB) complex defines a transcriptional program to promote the proliferation of cardiomyocytes. PLoS Genetics, 16(5), e1008818.
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3

Western Blotting of Cell Signaling Proteins

Cited 1 time
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Western blotting was conducted as previously described [26 (link)]. In Western blotting, the following antibodies were used at the indicated dilutions: p70 S6K–pT389 (1:1000, #9234), p70 S6K (1:1000, #9202), phospho–CK2 substrate (1:1000, #8738) from Cell Signaling Technology (Danvers, MA, USA); CK2β (1:500, 22418-1-AP), NOLC1 (1:1000, 11815-1-AP), and β-actin (1:5000, 66009-1-Ig) from Proteintech Group (Chicago, IL, USA); TOP2A-pS1469 (1:1000, PA5-64536) from Affinity Biosciences (Cincinnati, OH, USA); CK2β–pS209 (1:1000, 44-1090G) from Invitrogen (Waltham, MA, USA); TOP2A (1:200, sc-365916), CK2α (1:1000, sc-373894) from Santa Cruz Biotechnology (Dallas, TX, USA); goat anti-mouse IgG-HRP (1:5000, 115-035-062), goat anti-rabbit IgG-HRP (1:5000, 111-035-003) from Jackson ImmunoResearch Laboratories (West Grove, PA, USA).
Okanishi H., Ohgaki R., Xu M., Endou H, & Kanai Y. (2022). Phosphoproteomics revealed cellular signals immediately responding to disruption of cancer amino acid homeostasis induced by inhibition of l-type amino acid transporter 1. Cancer & Metabolism, 10, 18.
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4

Western Blot Profiling of Nuclear Factors

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Nuclear or Whole Cell Extracts were prepared, and protein concentrations were measured. 15ug of protein was run on a Bis-Tris gel, transferred at 200mA for 2 hours and blocked in 5% non-fat milk for 1 hour. Blots were incubated with primary antibodies overnight and then washed 3x in TBST. Blots were then incubated with secondary antibodies with conjugated −680nm and −800nm fluorophores (Licor, 1:25,000) for 45 minutes at room temperature, washed 3x, and then developed on an Odyssey® CLx Imaging System (Licor, Lincoln, NE United States). KDM4B/JMJD2B (Bethyl Laboratories A301–478A, 1:1000), TOP2A (Santa Cruz Biotech sc-3659, clone F-12, 1:1000), TOP2B (Santa Cruz Biotech H-286, sc-13059, 1:1000), MDR1/ABCB1 (Cell Signalling Technologies, 12683S, 1:1000), RNF2 (Cell Signalling Technologies, 5694S, 1:1000), H3 (Epicypher, SKU: 13–0001, 1:5000) We were unable to validate any of the available KAT6B antibodies. Licor software (Image Studio version 5.2.5) was used for the following: Image adjustment was limited to exposure (brightness and contrast; all lanes done equally and simultaneously) followed by cropping and exporting in .png format. No additional image manipulation was performed.
Seoane J.A., Kirkland J.G., Caswell-Jin J.L., Crabtree G.R, & Curtis C. (2019). Chromatin Regulators Mediate Anthracycline Sensitivity in Breast Cancer. Nature medicine, 25(11), 1721-1727.
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5

Detailed Antibody List for Neurological Studies

Cited 1 time
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Antibodies used in this study were the following: BrdU (1:500, AbDserotec, OBT0030CX), Brg1 (1:200, Santa Cruz Biotechnology, sc-374197), Cadps2 (1:500, Merck Millipore, ABN326), Calbindin (1:500, Swant, CB38), Chd7 (1:400, Cell Signaling, #6505), cleaved Caspase3 (1:200, Cell Signaling, #9661), Dab1 (1:200, Sigma-Aldrich, Ab232), Gap43 (1:500, Sigma-Aldrich, G9264), Ki67 (1:1,000, Abcam, ab15580), Ku70 (1:500, ThermoFisher Scientific, MA5-13110), GFP (1:500, Life technologies A11122; Abcam ab13970), H3K27Ac (2 μg per ChIP, Diagenode, pAb-174-050), NeuN (1:500, Merck Millipore, MAB377), p27 (1:500, BD bioscience, 610241), Parp (1:500, Cell Signaling,), Pax2 (1:1,000, Life technologies, 71-6000), Pax6 (1:500, Covance, PRB-278P), Reln (1:200, Merck Millipore, MAB5366), RPA116 (ref. 54 (link)), Sox2 (1:200, Santa Cruz Biotechnology, sc17320), Top2a (1:500, Santa Cruz Biotechnology, sc3659), Top2b (1:500, Santa Cruz Biotechnology, sc13059) and Zfpm2 (1:200, Santa Cruz Biotechnology, sc10755). These antibodies were quality-checked in previous publications.
Feng W., Kawauchi D., Körkel-Qu H., Deng H., Serger E., Sieber L., Lieberman J.A., Jimeno-González S., Lambo S., Hanna B.S., Harim Y., Jansen M., Neuerburg A., Friesen O., Zuckermann M., Rajendran V., Gronych J., Ayrault O., Korshunov A., Jones D.T., Kool M., Northcott P.A., Lichter P., Cortés-Ledesma F., Pfister S.M, & Liu H.K. (2017). Chd7 is indispensable for mammalian brain development through activation of a neuronal differentiation programme. Nature Communications, 8, 14758.
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