Ab226972

Manufactured by Abcam

Sourced in United Kingdom

Ab226972 is a laboratory equipment product offered by Abcam. It is a device designed for a specific function within the context of scientific research and laboratory work. The core function of this product is to perform a particular task or measurement, but a detailed description cannot be provided while maintaining an unbiased and factual approach.

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Lab products found in correlation

4 protocols using ab226972

Western Blot Analysis of FGF9 and CCND2

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For the western blot analyses, RIPA buffer containing protease inhibitors and phospha tase inhibitors (Roche, Basel, Switzerland) was used to prepare whole-cell lysates. Briefly, equal amounts of lysate were separated by SDS-polyacrylamide gel electrophoresis and then transferred to PVDF membranes (Millipore,Massachusetts, USA). After blocking with 5% bovine serum albumin, the membranes were probed with anti-FGF9 or CCND2 and anti-GAPDH (ab71395, ab226972, ab9485, Abcam, Cambridge, UK), followed by incubation with a horseradish peroxidase–conjugated secondary antibody [goat-anti-mouse IgG (1:2000) and goat-anti-rabbit IgG (1:3000)]. The proteins were visualized using Image Reader LAS-4000 (Fujifilm) and analyzed with Multi Gauge V3.2 software (GE Healthcare Life Sciences, USA).

He X., Chen S.Y., Yang Z., Zhang J., Wang W., Liu M.Y., Niu Y., Wei X.M., Li H.M., Hu W.N, & Sun G.G. (2018). miR-4317 suppresses non-small cell lung cancer (NSCLC) by targeting fibroblast growth factor 9 (FGF9) and cyclin D2 (CCND2). Journal of Experimental & Clinical Cancer Research : CR, 37, 230.

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Western Blotting for CCNE2 and GAPDH

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Western blotting was conducted according to a previous report (15 (link)) using antibodies against CCNE2 (ab226972, Abcam) and GAPDH (ab9485, Abcam).

Liu B., Qian D., Zhou W., Jiang H., Xiang Z, & Wu D. (2021). A Novel Androgen-Induced lncRNA FAM83H-AS1 Promotes Prostate Cancer Progression via the miR-15a/CCNE2 Axis. Frontiers in Oncology, 10, 620306.

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Western Blot Analysis of Bladder Cancer Cells

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First, protein was extracted by NP40 from bladder cancer cells, and it was separated on 10% SDS-PAGE gel and then transferred to polyvinylidene fluoride (PVDF) membranes (Bio-Rad). Nonspecific binding was blocked by incubating the PVDF membranes with 5% nonfat milk for 90 min. The membrane was then incubated with primary antibodies, including anti-CCND2 (1:1,000 dilution; ab226972, Abcam), anti-EZH2 (1:1,000 dilution; 21800-1-AP, Proteintech), anti-β-actin (1:2,000 dilution; 23660-1-AP, Proteintech), and anti-GAPDH (1:2,000 dilution; 10494-1-AP, Proteintech) in TBST solution at 4°C overnight. After washing with TBST, the PVDF membranes were incubated with horseradish peroxidase-conjugated secondary antibody (1:4,000 dilution; Boster Biological Technology) for 1.5 h at 37°C. At last the proteins were visualized using ECL-plus detection system (Pierce).

Yu G., Zhou H., Yao W., Meng L, & Lang B. (2019). lncRNA TUG1 Promotes Cisplatin Resistance by Regulating CCND2 via Epigenetically Silencing miR-194-5p in Bladder Cancer. Molecular Therapy. Nucleic Acids, 16, 257-271.

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BRCA Cell Lines and Antibody Analysis

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Human BRCA cell lines (MCF7, MDA-MB231) and an FBXL8-negative control breast cell line (MCF10A) were obtained from American Type Culture Collection (ATCC, Gaithersburg, Maryland, USA). MCF7 and MDA-MB231 were cultured in complete DMEM medium (Gibco, Paisley, Scotland, UK). MCF10A was cultured in complete DMEM/F12 medium (Gibco). All the complete media were supplemented with 10% FBS (Thermo Scientific) and 100 U/mL penicillin and 100 μg/mL streptomycin (Invitrogen, Carlsbad, CA, USA). Antibodies used in IHC (immunohistochemistry), immune blotting and immunoprecipitation analysis were anti-FBXL8 antibody (sc-390582, Santa Cruz Biotechnology, Dallas, TX, USA), anti-ubiquitin antibody (ab7780, Abcam, Cambridge, UK), anti-CCND2 antibody (ab226972, Abcam), anti-IRF5 antibody (ab2932, Abcam), normal mouse IgG2b (sc-3879, Santa Cruz Biotechnology) and anti-Tubulin antibody (ab176560, Abcam).

Chang S.C., Hsu W., Su E.C., Hung C.S, & Ding J.L. (2020). Human FBXL8 Is a Novel E3 Ligase Which Promotes BRCA Metastasis by Stimulating Pro-Tumorigenic Cytokines and Inhibiting Tumor Suppressors. Cancers, 12(8), 2210.

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