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Ab58023

Manufactured by Abcam
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Ab58023 is a primary antibody product manufactured by Abcam. It is designed for use in various immunoassay techniques.

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Lab products found in correlation

Apc 052, by Alomone (2 mentions) Alexa 488 conjugated goat anti rabbit secondary antibody, by Thermo Fisher Scientific (1 mentions) A21422, by Thermo Fisher Scientific (1 mentions) A11078, by Thermo Fisher Scientific (1 mentions) Sc 50446, by Santa Cruz Biotechnology (1 mentions) Sc 365685, by Santa Cruz Biotechnology (1 mentions) Ab21633, by Abcam (1 mentions) Jbw301, by Merck Group (1 mentions) Normal mouse igg, by Merck Group (1 mentions) Anti phospho histone h2a x ser139, by Merck Group (1 mentions) Nb200 109, by Novus Biologicals (1 mentions) Sc 53993, by Santa Cruz Biotechnology (1 mentions) Anti mouse igg hrp, by Cell Signaling Technology (1 mentions) Immobilon p pvdf membrane, by Merck Group (1 mentions) M11217, by Thermo Fisher Scientific (1 mentions) Western lighting plus ecl kit, by PerkinElmer (1 mentions) Donkey anti rabbit secondary antibody, by Jackson ImmunoResearch (1 mentions) Ma5 15256, by Thermo Fisher Scientific (1 mentions) Apc 030, by Alomone (1 mentions) Mini protean tgx gel, by Bio-Rad (1 mentions)

4 protocols using ab58023

1

Immunofluorescence Imaging of HCN4 and SGO1

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Cells were plated on 0.2% gelatin-coated coverslips and cultured for 3 days. Cultured cells were washed twice with 0.1 M phosphate-buffered saline (PBS) and fixed with paraformaldehyde (4%) in PBS for 10 min at room temperature, followed by several washes in PBS. Cells were subsequently incubated in PBS containing 2% normal goat serum for 30 min to block non-specific binding. They were then incubated with rabbit polyclonal anti-HCN4 antibody (1:200; APC-052, Alomone Labs) mixed with mouse monoclonal anti-SGO1 (1:200; ab58023, Abcam) in 2% normal goat serum PBS at 4 °C overnight. After washing with PBS, the cells were incubated with Alexa 488-conjugated goat anti-rabbit secondary antibody (1:1000; A-11078, Thermo Fisher Scientific) and an Alexa 555-conjugated goat anti-mouse secondary antibody (1:1000; A-21422, Thermo Fisher Scientific) for 1 h at room temperature. Nuclei were stained with 1 µM TOPRO3. Negative controls were performed by omitting the primary antibodies from the incubation solution. The fluorescence images were obtained with a Zeiss LSM‐710 inverted confocal laser scanning microscope (Carl Zeiss Meditec GmbH, Germany) and ZEN2.6 software was utilized for image collection.
Liu D., Song A.T., Qi X., van Vliet P.P., Xiao J., Xiong F., Andelfinger G, & Nattel S. (2021). Cohesin-protein Shugoshin-1 controls cardiac automaticity via HCN4 pacemaker channel. Nature Communications, 12, 2551.
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2

Western Blot Analysis of Cell Signaling Proteins

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Protein lysates were extracted from fibroblasts. A total of 25 μg of proteins were loaded on 12% sodium dodecyl sulfate–polyacrylamide gel electrophoresis (4561034; Bio-Rad, Hercules, CA) and transferred to a nitrocellulose membrane. Membranes were probed against SGO1 (mouse monoclonal, ab58023; Abcam, Cambridge, UK), BUB1 (mouse monoclonal, sc-365685; Santa Cruz, Dallas, TX), SM22α (rabbit polyclonal, sc-50446; Santa Cruz), and TPM1 (rabbit polyclonal, sc-28543; Santa Cruz) primary antibodies diluted 1:200 followed by horseradish-peroxidase–conjugated secondary antibodies. The blots were developed using SuperSignal (34580; Thermo Scientific). Signal intensity was quantified using ImageJ software (v.1.48; National Institutes of Health, Bethesda, MD).45 (link)
Piché J., Gosset N., Legault L.M., Pacis A., Oneglia A., Caron M., Chetaille P., Barreiro L., Liu D., Qi X., Nattel S., Leclerc S., Breton-Larrivée M., McGraw S, & Andelfinger G. (2018). Molecular Signature of CAID Syndrome: Noncanonical Roles of SGO1 in Regulation of TGF-β Signaling and Epigenomics. Cellular and Molecular Gastroenterology and Hepatology, 7(2), 411-431.
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3

Antibody Panel for Protein Analysis

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The following antibodies were used: anti-SGO1 mouse monoclonal (ab58023, Abcam); anti-SGO1 rabbit polyclonal (GTX117103, GeneTex, and ab21633, Abcam); anti-MYCN mouse monoclonals OP13 (Calbiochem), NB200-109 (Novus Biologicals), and sc-53993 (Santa Cruz Biotechnology); anti-MYCN rabbit polyclonal (#9405, Cell Signaling Technology); anti-phospho-histone H2A.X (Ser139) mouse monoclonal (JBW301, Millipore); anti-SMC3 rabbit polyclonal (A300-060A, Bethyl laboratories); anti-β-ACTIN (8H10D10) mouse monoclonal (#3700, Cell Signaling Technology); normal mouse IgG (#12–371, Millipore); goat anti-mouse IgG (H+L) secondary antibody, Alexa Fluor® 488 conjugate (A-11001, Thermo Fisher Scientific); anti-mouse IgG, HRP-linked (#7076S, Cell Signaling Technology); and anti-rabbit IgG, HRP-linked (#7074S, Cell Signaling Technology) antibodies.
Murakami-Tonami Y., Ikeda H., Yamagishi R., Inayoshi M., Inagaki S., Kishida S., Komata Y., Jan Koster J.K., Takeuchi I., Kondo Y., Maeda T., Sekido Y., Murakami H, & Kadomatsu K. (2016). SGO1 is involved in the DNA damage response in MYCN-amplified neuroblastoma cells. Scientific Reports, 6, 31615.
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4

Western Blot Protein Analysis

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Western blot analysis was performed on Mini-PROTEAN® TGX™ Gels (Bio-Rad Laboratories). Proteins were electrotransferred onto Immobilon-P PVDF membrane (millipore, pore size 0.45 mm). After blocking 1 h in Tris-HCl-buffered saline with 0.2% Tween-20 (TBST) and 5% non-fat milk (Bioshop® Canada), the membranes were incubated overnight with primary antibody in TBST containing 2% non-fat milk. The rabbit polyclonal anti-HCN2 antibody (1:500; APC-030, Alomone Labs), rabbit polyclonal anti-HCN4 antibody (1:1000; APC-052, Alomone Labs), mouse monoclonal anti-SGO1 antibody (1:1000; ab58023, Abcam), mouse monoclonal anti-GAPDH antibody (1:1000; 10R-G109a, Fitzgerald), mouse monoclonal anti-GFP antibody (1:1000; MA5-15256, Invitrogen), rat monoclonal anti-mCherry (1:1000; M11217, Invitrogen) and mouse monoclonal anti-PP2A C subunit (1:1000; SAB4200266, Sigma-Aldrich) were utilized to detect HCN2, HCN4, SGO1, GAPDH, GFP, mCherry and PP2A C proteins respectively. Detection was carried out by the use of horseradish peroxidase conjugated secondary antibodies (Donkey anti rabbit secondary antibody, 1:5000, #711035152, Jackson; Donkey anti mouse secondary antibody, 1:5000, #711035151, Jackson; Donkey anti rat secondary antibody, 1:5000, #A18745, Invitrogen), photographic film (Agfa NV) and the Western Lighting® plus ECL kit (PerkinElmer).
Liu D., Song A.T., Qi X., van Vliet P.P., Xiao J., Xiong F., Andelfinger G, & Nattel S. (2021). Cohesin-protein Shugoshin-1 controls cardiac automaticity via HCN4 pacemaker channel. Nature Communications, 12, 2551.
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