Cyclin d1 sc 718

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Cyclin D1 (sc-718) is a primary antibody produced by Santa Cruz Biotechnology. It is designed to detect Cyclin D1 protein, which is a key regulator of the cell cycle progression from G1 to S phase. The antibody can be used in various immunoassay applications to study the expression and localization of Cyclin D1 in different cell types and tissues.

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4 protocols using cyclin d1 sc 718

Protein Extraction and Western Blot Analysis

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Cells and tumor tissues were lysed with radioimmunoprecipitation assay buffer. After incubation, cell extracts were centrifuged and the supernatant was collected for Bradford assay to quantify protein concentration by a BioTek Synergy HTX Multi‐Mode Microplate Reader (BioTek, Winooski, VT). Equal amounts (20–50 µg) of protein were loaded for sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS‐PAGE). The proteins inside the gels were transferred onto nitrocellulose membrane with a wet transfer method. The membranes were blocked with 3% milk for 1 h. Primary antibodies were incubated overnight. Secondary antibodies were incubated for 1 h. Antibodies for ferritin heavy chain (FTH1, #3998), active beta catenin (#8814), Anti‐rabbit IgG HRP‐linked Antibody (#7074), and Anti‐mouse IgG HRP‐linked Antibody (#7076) were from Cell signaling Technology (Danvers, MA). Primary antibodies for TFRC (sc‐393719), HIF‐1α (sc‐13515), HIF‐2α (sc‐13596), pS6 (sc‐514033), pS6K (sc‐8418), p‐p53 (sc‐377567), p53 (sc‐6243), c‐MYC (sc‐40), GAPDH (sc‐47724), Cyclin D1 (sc‐718), and Actin (sc‐8432) were from Santa Cruz Biotechnology (Dallas, TX). P‐RPA2 (PA5‐39809) was from Invitrogen (Carlsbad, CA). POLD1 (15646‐1‐AP), p‐CHK1 (28805‐1‐AP), E2F1 (66515‐1‐Ig), Axin2 (20540‐1‐AP), and TNKS (18030‐1‐AP) were from Proteintech (Rosemont, IL).

Kim H., Villareal L.B., Liu Z., Haneef M., Falcon D.M., Martin D.R., Lee H., Dame M.K., Attili D., Chen Y., Varani J., Spence J.R., Kovbasnjuk O., Colacino J.A., Lyssiotis C.A., Lin H.C., Shah Y.M, & Xue X. (2023). Transferrin Receptor‐Mediated Iron Uptake Promotes Colon Tumorigenesis. Advanced Science, 10(10), 2207693.

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Butein and Cisplatin Signaling Pathways

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Butein and cisplatin (CDDP) were obtained from Sigma (St. Louis, MO, USA), and were dissolved in dimethyl sulfoxide (DMSO) and stored at −20°C until use. The ERK inhibitor, U0126, p38 inhibitor, SB203580, and the AKT inhibitor, LY294002, were obtained from Sigma and used at final concentrations of 10, 20 and 20 µM, respectively. The following antibodies were used in western blot analysis: anti-ERK1/2 (#9102; 1:1,000), anti-p-ERK1/2 (Thr202/Tyr204) (#9101; 1:1,000), anti-p38 (#9212; 1:1,000), anti-p-p38 (Thr180/Tyr182) (#9211; 1:1,000), anti-Akt (#9272; 1:1,000) and anti-p-Akt (Ser413) (#9271; 1:1,000) (all purchased from Cell Signaling Technology, Danvers, MA, USA). β-actin (sc-4778; 1:5,000), forkhead box O3a (FoxO3 or FoxO3a; sc-9812; 1:1,000), Bax (sc-7480; 1:1,000), Bcl-2 (sc-7382; 1:1,000), p27 (sc-1641; 1:1,000) and cyclin D1 (sc-718; 1:1,000) antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Horseradish peroxidase (HRP)-conjugated goat anti-mouse/anti-rabbit immunoglobulin (IgG; ab6721; 1:1,000) was obtained from Abcam (Hong Kong, China).

ZHANG L., YANG X., LI X., LI C., ZHAO L., ZHOU Y, & HOU H. (2015). Butein sensitizes HeLa cells to cisplatin through the AKT and ERK/p38 MAPK pathways by targeting FoxO3a. International Journal of Molecular Medicine, 36(4), 957-966.

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Apoptosis and Metastasis Signaling Pathway

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The following antibodies and reagents were purchased: Phospho-Stat Antibody Sampler Kit # 9914T, Death Receptor Antibody Sampler Kit #8356, Caspase-3 #9662S, cleaved caspase‐3 #9664, MMP-9 #3852S, and Survivin #2808S (Cell Signaling Technology, Danvers, USA); Anti-MAGEC2 antibody #ab209667 (Abcam, Cambridge, USA); cyclin D1 #sc-718, BID #sc-11423, β‐actin #sc‐58673 (Santa Cruz Biotechnology, Santa Cruz, USA); PARP #51-6639GR (BD Biosciences, Franklin Lakes, USA); Stattic #S7947 (Sigma-Aldrich, St. Louis, USA).

Park D., Han S., Joo H., Ka H.I., Soh S., Park J, & Yang Y. (2021). Increased Melanoma-Associated Antigen C2 Expression Affords Resistance to Apoptotic Deathin Suspension-Cultured Tumor Cells. Journal of Breast Cancer, 24(2), 138-152.

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Cell Signaling Protein Regulation

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Human recombinant VEGF protein was purchased from R&D Systems (Minneapolis, MN). Rabbit anti-human c-fos (sc-52), c-jun (sc-1694), and cyclin D1 (sc-718) antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Rabbit anti-human JunB (Cat. No. 3755) and JunD (Cat. No. 5000) were purchased from Cell Signaling (Danvers, MA). Mouse monoclonal anti-Flag antibody was from Sigma. Actinomycin D were purchased from EMD Millipore (Billerica, MA)

Jia J., Ye T., Cui P., Hua Q., Zeng H, & Zhao D. (2016). AP-1 transcription factor mediates VEGF-induced endothelial cell migration and proliferation. Microvascular research, 105, 103-108.

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