Sc 22760

Manufactured by Abcam

Sc-22760 is a primary antibody developed by Santa Cruz Biotechnology. It is designed for the detection of FOXO1 protein expression in various sample types.

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Lab products found in correlation

Immunofluorescence microscope, by Zeiss (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by Dojindo Laboratories (2 mentions) Ab36823, by Abcam (1 mentions) Ab6161, by Abcam (1 mentions) Slowfade, by Thermo Fisher Scientific (1 mentions) Click it reaction buffer, by Thermo Fisher Scientific (1 mentions) γh2ax, by Merck Group (1 mentions) Ab36823, by Proteintech (1 mentions) Ab6672, by Abcam (1 mentions) Ab16048, by Santa Cruz Biotechnology (1 mentions)

4 protocols using sc 22760

Immunofluorescence Analysis of DNA Damage Markers

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Immunofluorescence analysis was performed using antibodies against γ-H2AX (1:1,000, Millipore, 05-636), phosphor-(Ser/Thr) ATM/ATR substrate (1:500, Cell Signaling Technology, 2851) and 53BP1 (1:1,000, Santa Cruz, sc-22760; 1:1,000, abcam, ab36823). DNA was stained with 2 mg ml⁻¹ 4′,6-diamidino-2-phenylindole (Dojindo). Fluorescence images were observed and photographed using an immunofluorescence microscope (Carl Zeiss)14 (link)62 (link).

Takahashi A., Okada R., Nagao K., Kawamata Y., Hanyu A., Yoshimoto S., Takasugi M., Watanabe S., Kanemaki M.T., Obuse C, & Hara E. (2017). Exosomes maintain cellular homeostasis by excreting harmful DNA from cells. Nature Communications, 8, 15287.

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Immunoblotting for 53BP1 and Tubulin

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Immunoblotting was performed with total cellular extract using standard protocols. Antibodies used were rabbit anti-53BP1 (Santa-Cruz, sc-22760) and rat anti-tubulin (Abcam, ab6161).

Binan L., Bélanger F., Uriarte M., Lemay J.F., Pelletier De Koninck J.C., Roy J., Affar E.B., Drobetsky E., Wurtele H, & Costantino S. (2019). Opto-magnetic capture of individual cells based on visual phenotypes. eLife, 8, e45239.

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Immunofluorescence Staining of DNA Damage Markers

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After incubation with pneumolysin, cells were permeabilized with 0.2% Triton X-100 in PBS for 10 min, and blocked with 3% bovine serum albumin (BSA) in PBS for 40 min. Primary antibodies against γH2AX (Ser-139) (Millipore #05–636), 53BP1 (Santa Cruz #sc-22760), MDC1 (Abcam #1169), p53BP1 (S1778, Cell signaling #2675) were used at 1:100 dilutions in PBS, and incubated for 1 h at room temperature with cover slips. For TUNEL staining, the labeling enzyme (Roche #1 1684795 910) was incubated similarly for 1 h at 37 °C. Secondary antibodies (Invitrogen) labeled with either Alexa Fluor 488 or 564 or 647 were used. For EdU detection, Alexa Fluor azide was used in Click-iT reaction buffer (Invitrogen). Slides were mounted with SlowFade (Invitrogen) and stored at 4 °C until imaging. All stained slides were examined under confocal microscope, and 15 images (at 5 × 3 sites) were taken for each well under 40X magnification.

Rai P., He F., Kwang J., Engelward B.P, & Chow V.T. (2016). Pneumococcal Pneumolysin Induces DNA Damage and Cell Cycle Arrest. Scientific Reports, 6, 22972.

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Immunofluorescence Analysis of Cellular Senescence

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Immunofluorescence analysis was performed using antibodies against Lamin B1 (Abcam, ab16048, 1:1000 dilution), dsDNA marker (Santa Cruz, sc-58749, 1:250 dilution), γ-H2AX (Millipore, 05-636, 1:2000 dilution), phosphor-(Ser/Thr) ATM/ATR substrate (Cell Signaling Technology, 2851, 1:500 dilution), p16 (Santa Cruz, sc-468, 1:500 dilution), p21 (Abcam, ab2961, 1:50 dilution), 53BP1 (Santa Cruz, sc-22760; Abcam, ab36823, 1:500 dilution), IL-1β (Proteintech, 16806-1-AP, 1:300 dilution), IL-6 (Abcam, ab6672, 1:400 dilution), STING (Abcam, ab92605, 1:200 dilution), phospho-TBK1 (Bioss antibodies, bs-3440R, 1:100 dilution), IFN-β (Abcam, ab140211, 1:100 dilution), DNase2 (Bioss antibodies, bs-7652, 1:100 dilution), TREX1 (Novus Biologicals, NBP1-76977, 1:100 dilution) and Desmin (Thermo Fisher Scientific, MA5-13259, 1:100 dilution). DNA was stained with DAPI (Dojindo). Fluorescence images were observed and photographed using an immunofluorescence microscope (Carl Zeiss AG)^{11 (link),43 (link)}.

Takahashi A., Loo T.M., Okada R., Kamachi F., Watanabe Y., Wakita M., Watanabe S., Kawamoto S., Miyata K., Barber G.N., Ohtani N, & Hara E. (2018). Downregulation of cytoplasmic DNases is implicated in cytoplasmic DNA accumulation and SASP in senescent cells. Nature Communications, 9, 1249.

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