Chemiluminescent immunoblotting reagent

Manufactured by Thermo Fisher Scientific

Sourced in United States

Chemiluminescent immunoblotting reagent is a laboratory product used to detect and quantify specific proteins within a sample. It facilitates the visualization of protein targets in Western blotting applications.

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Chemiluminescent reagent

4 protocols using chemiluminescent immunoblotting reagent

Curcumin Analog B14 Inhibits Breast Cancer

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Curcumin was purchased from Sigma (St. Louis, MO). B14, a Curcumin analog (Figure 1), was provided by Dr. Ge RS (Department of Anesthesiology, the Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University).
In all in vitro experiments, Curcumin and B14 compounds were dissolved in medium containing 0.1% DMSO. Human breast cancer cell lines MCF‐7 and MDA‐MB‐231 were purchased from Shanghai Institute of Life Sciences Cell Resource Center (Shanghai, China). DMEM and FBS were obtained from Gibco BRL (Grand Island, NY, USA). MTT reagent was from Sigma (St. Louis, MO). The FITC Annexin V Apoptosis Detection Kit and Cell Cycle Staining Kit were purchased from BD Pharmingen (Franklin Lakes, NJ). GAPDH, Cyclin E1, Cyclin D1, Bax, Bcl‐2, Cyto‐C, TIMP1, Caspase3, cleaved Caspase3, and cleaved Caspase7 antibodies were purchased from Abcam company. Modified RIPA lysis buffer, protease inhibitor cocktail and chemiluminescent immunoblotting reagent were obtained from Thermo Fisher Scientific (Rockford, IL, USA).

Shen H., Shen J., Pan H., Xu L., Sheng H., Liu B, & Yao M. (2020). Curcumin analog B14 has high bioavailability and enhances the effect of anti‐breast cancer cells in vitro and in vivo. Cancer Science, 112(2), 815-827.

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Molecular Mechanisms of NLRP3 Inflammasome Activation

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Dulbecco’s Modified Eagle Medium (DMEM) was purchased from Gibco (Grand Island, NY, USA). Fetal bovine serum (FBS) was purchased from Thermo Scientific (Waltham, MA, USA). Modified radioimmunoprecipitation assay (RIPA) lysis buffer, protease inhibitor cocktail, Coomassie Plus™ Protein Assay Reagent, and chemiluminescent immunoblotting reagent were obtained from Thermo Fisher Scientific (Rockford, IL, USA). QIAzol lysis reagent was purchased from Qiagen (Valencia, CA, USA). ReverTra Ace^® qPCR Master Mix was purchased from Toyobo Co., Ltd. (Osaka, Japan). SensiFAST^TM SYBR^® Lo-ROX Kit was purchased from Meridian Bioscience^® (Cincinnati, OH, USA). The MTT or 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide dye and mouse anti-β-actin primary antibody were purchased from Sigma-Aldrich Company (St. Louis, MO, USA). Anti-NLRP3 primary antibody, anti-ASC primary antibody, anti-caspase-1 primary antibody, anti-NF-κB primary antibody, anti-PARP primary antibody, and goat horseradish peroxidase-conjugated anti-mouse- or anti-rabbit-IgG were obtained from Cell Signaling Technology company (Danvers, MA, USA).

Semmarath W., Srisawad K., Arjsri P., Umsumarng S., Yodkeeree S., Jamjod S., Prom-u-thai C, & Dejkriengkraikul P. (2023). Protective Effects of Proanthocyanidin-Rich Fraction from Red Rice Germ and Bran on Lung Cell Inflammation via Inhibition of NF-κB/NLRP3 Inflammasome Pathway. Nutrients, 15(17), 3793.

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Protein Expression Analysis Protocol

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Fetal bovine serum and RPMI‐1640 were purchased from Gibco BRL Company (Grand Island, NY, USA). Gelatinase A was purchased from Sigma Chemical Company (St Louis, MO, USA). Modified RIPA lysis buffer, protease inhibitor cocktail and chemiluminescent immunoblotting reagent were obtained from Thermo Fisher Scientific (Rockford, IL, USA).

Mapoung S., Suzuki S., Fuji S., Naiki‐Ito A., Kato H., Yodkeeree S., Ovatlarnporn C., Takahashi S, & Limtrakul (Dejkriengkraikul) P. (2018). Cyclohexanone curcumin analogs inhibit the progression of castration‐resistant prostate cancer in vitro and in vivo. Cancer Science, 110(2), 596-607.

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Molecular Mechanisms of Phytochemical-Induced Effects

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Fetal bovine serum (FBS), Roswell Park Memorial Institute (RPMI)-1640 and penicillin-streptomycin were purchased from Gibco BRL Company (Grand Island, NY, USA). Modified Radioimmunoprecipitation assay (RIPA) lysis buffer, protease inhibitor cocktail, Commasie Plus™ Protein Assay Reagent, and chemiluminescent immunoblotting reagent were obtained from Thermo Fisher Scientific (Rockford, IL, USA). Syringic acid, pyrogallol, p-coumaric acid, catechin, gallic acid, protocatechuic acid, chlorogenic acid, caffeic acid, vanillic acid, ferulic acid, and ellagic acid, Sulforhodamine B (SRB) reagent, mitochondria membrane potential kit (JC-10 dye), propidium iodide (PI) and anti-β-actin antibody were obtained from Sigma-Aldrich (St. Louis, MO, USA). Primary antibody of PARP-1 was purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Primary antibodies of cyclin D1, cyclin E, CDK-4, CDK-6, cleaved-caspase3, phospho-GSK-3β, GSK-3β (Ser9), phospho-β-catenin, β-catenin, phospho-Akt, Akt, and horseradish peroxidase-conjugated anti-mouse or rabbit-IgG were purchased from Cell Signaling Technology (Beverly, MA, USA). Apoptosis assay kits were obtained from Bio-Legend (San Diego, CA, USA).

Arjsri P., Mapoung S., Semmarath W., Srisawad K., Tuntiwechapikul W., Yodkeeree S, & Dejkriengkraikul P. (2023). Pyrogallol from Spirogyra neglecta Inhibits Proliferation and Promotes Apoptosis in Castration-Resistant Prostate Cancer Cells via Modulating Akt/GSK-3β/β-catenin Signaling Pathway. International Journal of Molecular Sciences, 24(7), 6452.

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