Anti cdc20

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Anti-Cdc20 is a lab equipment product that functions as an antibody targeting the Cdc20 protein. Cdc20 is a key regulator of the cell cycle and is involved in the activation of the anaphase-promoting complex/cyclosome (APC/C). The Anti-Cdc20 product can be used in research applications to study the role of Cdc20 in cellular processes.

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Lab products found in correlation

7 protocols using anti cdc20

Generation and Characterization of Anti-BEX4 Antibodies

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Mouse polyclonal antibodies against human BEX4 protein were generated in C57BL/6 mice. Briefly, purified GST-BEX4 protein was injected four times intraperitoneally. Rabbit polyclonal antibodies against C-terminal polypeptides 89–106 of human BEX4 protein were commercially generated (Youngin Frontier, Seoul, Korea). The other antibodies used in this study were as follows: anti-GFP, anti-PLK1, anti-CDK1, anti-aurora A, anti-cIAP-1, anti-cdc20 (Santa Cruz Biotechnology, Dallas, TX, USA), anti-actin (Sigma-Aldrich, St. Louis, MO, USA), anti-poly(ADP-ribose) polymerase-1 (PARP), anti-cleaved-caspase 9, cleaved-caspase 7, anti-active-caspase 3, anti-phospho-threonine (p-Thr) (Cell Signaling Technology, Danvers, MA, USA), anti-aurora B (BD Biosciences PharMingen, San Diego, CA, USA), anti-securin (PTTG1; Zymed, San Francisco, CA, USA), anti-Myc (Bethyl Laboratories, Montgomery, TX, USA), and Alexa Fluor (Invitrogen, Leek, The Netherlands). The following reagents were used: MG132, cycloheximide (CHX), dimethyl sulfoxide (DMSO) (AG Scientific, San Diego, CA, USA), nocodazole (Sigma-Aldrich), and PLK1 kinase inhibitor BI2536 (Axon Medchem, Groningen, Netherlands).

Lee J.K., Ha G.H., Kim H.S, & Lee C.W. (2018). Oncogenic potential of BEX4 is conferred by Polo-like kinase 1-mediated phosphorylation. Experimental & Molecular Medicine, 50(10), 138.

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Comprehensive Immunoblotting Antibody Panel

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The following commercially available primary antibodies were used for immunoblotting: anti-IRS2 (Cell Signaling Technologies, 4502) 1:750; anti-Cdh1/Fzr1 (Sigma Aldrich, CC43) 1:500; anti-anillin (a gift from Christine Field (21 (link))) 1:1000; anti-Aurora B (Bethyl, A300-431) 1:1000; anti-Eg5 (Cell Signaling Technologies, 4203) 1:1000; anti-Top2A (Cell Signaling Technologies, 12286) 1:1000; anti-TK1 (Cell Signaling Technologies, 8960) 1:1000; anti-Mps1 (Abcam, ab11108), 1:1000); anti-APC3 (BD Transduction Laboratories, 610455) 1:500; anti-cyclin B1 (Santa Cruz Biotechnology, sc-752) 1:500; anti-Cdc20 (Santa Cruz Biotechnology, sc-8358) 1:500; anti-c-Myc (9E10, Santa Cruz Biotechnology, sc-40) 1:1000; anti-HA-peroxidase (Sigma Aldrich), 1:1500; anti-cyclin A2 (Santa Cruz Biotechnology, sc-596) 1:500; anti-IRS1 (Cell Signaling Technologies, 2382) 1:750; anti-MyoD1 (Cell Signaling Technologies, 13812) 1:750; anti-GAPDH (Abcam, ab8245) 1:2000; anti-α tubulin (Abcam, ab7291 and Santa Cruz Biotechnology, sc-8035) 1:1000 for both; anti-vinculin (Santa Cruz Biotechnology, sc-73614) 1:2000. Secondary antibodies used: anti-rabbit IgG-HRP (GE Healthcare, NA934) and anti-mouse IgG-HRP (GE Healthcare, NA931V), both at 1:3000 dilutions.

Manohar S., Yu Q., Gygi S.P, & King R.W. (2020). The Insulin Receptor Adaptor IRS2 is an APC/C Substrate That Promotes Cell Cycle Protein Expression and a Robust Spindle Assembly Checkpoint. Molecular & Cellular Proteomics : MCP, 19(9), 1450-1467.

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Antibody Characterization for Cell Signaling

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The following antibodies were used for immunoblotting and immunoprecipitation: anti-Tubulin (Sigma, T9026), anti-BubR1 [BD Biosciences; Clone 9/BUBR1 (RUO)], anti-Bub3 [mouse, BD Biosciences; Clone 31/Bub3 (RUO)], anti-Bub3 (rabbit, Sigma, B7811), anti-GAPDH (Cell Signaling, 14C10), anti-GFP (Roche), anti-Myc (Roche), and anti-Cdc20 (goat, Santa Cruz Biotechnology). Fluorescent secondary antibodies were purchased from Life Technologies. Antibodies against Mad2, APC3, Mad1, Cdc20 (pS153), and Bub1 were previously described (Fang et al., 1998 (link); Kim et al., 2012 (link); Lin et al., 2014 (link); Tang et al., 2001 (link)). The Bub1 pS459 and pSpT antibodies and the Mad1 pT716 antibody were generated at an on-campus facility. Two Bub1 phospho-peptides, with the sequences of CKVQP[pS]PTVH and CKVQP[pS]P[pT]VHTK, and a Mad1 phospho-peptide, with the sequence of CELFSRQ[pT]VA, were used to immunize rabbits. The antibodies were affinity-purified with SulfoLink resins (Thermo Scientific) coupled to the phospho-peptides.

Ji Z., Gao H., Jia L., Li B, & Yu H. (2017). A sequential multi-target Mps1 phosphorylation cascade promotes spindle checkpoint signaling. eLife, 6, e22513.

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Comprehensive Protein Analysis Protocol

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Anti-DDB1, Bethyl (IHC-0013401), IHC (1:400), IF (1:200); Anti-DDB1, Epitomics (3821-1), WB (1:10,000); Anti-DCAF1, Proteintech (11612-1-AP), IHC (1:200), IF (1:200), WB (1:1,000); Anti-MYC, Cell Signaling (2272), WB (1:1,000); Anti-HA, Cell Signaling (3724), WB (1:1,000); Anti-HA, Sigma (F3165), WB (1:1,000); Anti-ERK1/2, Santa Cruz (sc-94), WB (1:1,000); anti-FITC-α-tubulin, Sigma (F2168), IF(1:500); anti-CREST, Fitzgerald Industries International (90C-CS1058), IF (1:100); anti-TOP2B, Epitomics (3747-1), IF (1:200); anti-SMC3, Abcam (ab128919), IF (1:20); anti-MAD2, Abcam (ab9777), IF (1:100); anti-PP2A-A, Cell Signaling (2041), WB (1:1,000), IF (1:100); anti-PP2A-B, Cell Signaling (2290), WB (1:1,000); anti-PP2A-C, Cell Signaling (2259), WB (1:1,000); anti-NEDD8, Epitomics (1571-1), WB (1:1,000); anti-cyclin B, Cell Signaling (4138), WB (1:1,000); anti-securin, Abcam (ab3305), WB (1:1000); anti-CDC20, Santa Cruz (sc-8358), WB (1:500); anti-pCDK1(T161), Cell Signaling (9114), WB (1:1,000); anti-pERK1/2, Cell Signaling (9101), WB (1:1,000).

Yu C., Ji S.Y., Sha Q.Q., Sun Q.Y, & Fan H.Y. (2015). CRL4–DCAF1 ubiquitin E3 ligase directs protein phosphatase 2A degradation to control oocyte meiotic maturation. Nature Communications, 6, 8017.

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Mitotic Checkpoint Protein Localization

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The antibodies used were anti-MAD1 (proteintech, #18322-1-AP; Santa Cruz, #sc-47746), anti-CENP-B (Santa Cruz, #sc-376283; #sc-22788), anti CDC20 (Santa Cruz, #sc-5296), anti-MAD2 (proteintech, #10337-1-AP; Covance, #PRB-452C-200), anti-BUBR1 (proteintech, #11504-2-AP), anti-cyclinB1 (abclonal, #A2056), anti-ZW10 (abcam, #ab21582), anti-Flag-tag (Sigma-Aldrich, #F1804), anti-ULK1 (Santa Cruz, #sc-33182; proteintech, #20986-1-AP), anti-Atg13 (CST, #13468S), anti-FIP200 (proteintech, #17250-1-AP), anti-Atg3 (MBL, #M133-3), anti-GFP-tag (Santa Cruz, #sc-9996), anti-His-tag (MBL, #D291-3), anti-BUB1 (proteintech, #13330-1-AP), anti-BUB3 (proteintech, #27073-1-AP), anti-H3S10 (abcam, #ab32017), anti-Knl1 (abclonal, A13108), anti-ROD (abclonal, A13064), anti-ZWILCH (proteintech, #14281-1-AP), anti-α-tubulin (CST, #3873T) and anti-actin (Santa Cruz, #sc-7210). A phospho (ph)-MAD1-S546 specific antibody was generated by Beijing Biodragon Immunotechnologies Co. Ltd.
Treatments included thymidine (Sigma, T9250), RO3306 (Selleck, S7747), Taxol (Santa Cruz, sc-201439), nocodazole (Selleck, S2775), vinblastine (Selleck, S4505).

Yuan F., Jin X., Li D., Song Y., Zhang N., Yang X., Wang L., Zhu W.G., Tian C, & Zhao Y. (2019). ULK1 phosphorylates Mad1 to regulate spindle assembly checkpoint. Nucleic Acids Research, 47(15), 8096-8110.

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Depletion of Cell Cycle Regulators

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For siRNA-mediated depletion, Cal51 cells were reverse transfected with 25 nM siRNA to the indicated genes using DharmaFECT1 (Dharmafect) transfection reagent. Transfected cells were harvested 54 hours later for RNA extraction, western blotting and fluorescence-activated cell sorting (FACS). Efficiency of depletion was either monitored by western blotting with the indicated antibodies or by qRT-PCR against the indicated genes. Antibodies used were anti-PRPF8 (Abcam), anti-Separase (Abcam), anti-CDC20 (Santa Cruz), anti-APC8 (Novus Biologicals), anti-Actin (Abcam).

Wickramasinghe V.O., Gonzàlez-Porta M., Perera D., Bartolozzi A.R., Sibley C.R., Hallegger M., Ule J., Marioni J.C, & Venkitaraman A.R. (2015). Regulation of constitutive and alternative mRNA splicing across the human transcriptome by PRPF8 is determined by 5′ splice site strength. Genome Biology, 16(1), 201.

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Western Blot Protein Analysis

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Western blot analyses were performed as previously described [40 (link)] using anti-cyclin (Santa Cruz Biotechnology), anti-Cdc27 (MBL International Corporation), anti-Cdc20 (Santa Cruz Biotechnology) anti-Cdh1 (Calbiochem), anti-Actin (Sigma-Aldrich), and anti-alpha-Tubulin (Sigma-Aldrich) as primary antibodies. The horseradish-peroxidase-conjugated secondary antibodies used were anti-rabbit (Amersham Biosciences) and anti-mouse (Sigma-Aldrich). Densitometric quantification was performed using ImageJ.

Schnerch D., Schüler J., Follo M., Felthaus J., Wider D., Klingner K., Greil C., Duyster J., Engelhardt M, & Wäsch R. (2017). Proteasome inhibition enhances the efficacy of volasertib-induced mitotic arrest in AML in vitro and prolongs survival in vivo. Oncotarget, 8(13), 21153-21166.

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