Ecl plus solution

Manufactured by Thermo Fisher Scientific

Sourced in United States

ECL Plus solution is a chemiluminescent detection reagent for Western blotting applications. It is designed to detect horseradish peroxidase (HRP)-labeled proteins on membrane blots.

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Lab products found in correlation

Goat anti mouse and goat anti rabbit hrp conjugated igg, by Bio-Rad (2 mentions) Anti nampt, by Fortis Life Sciences (1 mentions) Mtt solution, by Merck Group (1 mentions) C myc, by Cell Signaling Technology (1 mentions) Mouse monoclonal anti β actin, by Merck Group (1 mentions) β actin, by Cell Signaling Technology (1 mentions) β catenin, by Cell Signaling Technology (1 mentions) P pak4, by Cell Signaling Technology (1 mentions) Sirtuin 1, by Cell Signaling Technology (1 mentions) Fk866, by Bio-Techne (1 mentions) P β catenin, by Cell Signaling Technology (1 mentions) Sunitinib, by LC Laboratories (1 mentions) Parp antibody, by Cell Signaling Technology (1 mentions) Nrf2 antibody, by Abcam (1 mentions) Bptes, by Merck Group (1 mentions) Immobilon pvdf, by Merck Group (1 mentions) Anti akr1c1, by Abnova (1 mentions) Rabbit anti txnrd1, by Proteintech (1 mentions) Chemidoc xr, by Bio-Rad (1 mentions) Anti actin, by Merck Group (1 mentions)

Spelling variants of this product

ECL Plus (212 citations) ECL solution (151 citations) PLUSTM (3 citations) Pierce ECL Plus solution (2 citations) Pierce ECL Plus substrate solution (2 citations)

4 protocols using ecl plus solution

Protein Expression Quantification Protocol

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MTT solution, mouse monoclonal anti-β-actin, NA, NMN and NAD were obtained from Sigma (St. Louis, MO, USA). The antibodies against (PAK4, p-PAK4, β-catenin, p-β-catenin, cycline D1, c-Myc, β-actin (rabbit), PARP, Sirtuin 1) were from cell signaling Technology, Inc. (Beverly, MA, USA). Goat anti-mouse and goat anti-rabbit HRP conjugated IgG were obtained from Bio-Rad (Hercules, CA). Anti-NAMPT was from Bethyl Laboratories (Montgomery, TX, USA), anti-NAPRT1 was from Proteintech (Rosemont, IL 60018, USA). ECL Plus solution was from Thermo-Fisher Scientific (Waltham MA, USA). KPT-9274 and its vehicle were from Karyopharm Therapeutics (Newton, MA, USA). FK866 was from TOCRIS Biosciences. Sunitinib was obtained from LC laboratories (Woburn, MA).

Aboud O.A., Chen C.H., Senapedis W., Baloglu E., Argueta C, & Weiss R.H. (2016). Dual and specific inhibition of NAMPT and PAK4 by KPT-9274 decreases kidney cancer growth. Molecular cancer therapeutics, 15(9), 2119-2129.

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Antibody Characterization and Inhibitor Preparation

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PARP antibody was purchased from Cell Signaling Technology, Inc. (Beverly, MA, USA), goat anti-mouse and goat anti-rabbit HRP-conjugated IgG from Bio-Rad (Hercules, CA), anti-nuclear factor erythroid related factor 2 (NRF2) antibody from Abcam (Cambridge, MA) and ECL Plus solution from Thermo-Fisher Scientific (Waltham MA, USA). CB-839 was supplied by Calithera, Inc. (South San Francisco, CA) and was dissolved either in DMSO for in vitro experiments (10 mM stock, stored at -20 ⁰C) or in vehicle for in vivo experiments. The in vivo vehicle consisted of 25% (w/v) hydroxypropyl-β-cyclodextrin in 10 mmol/L citrate (pH 2). The GLS inhibitor BPTES (bis-2-(5-phenylacetamido-1,2,4-thiadiazol-2-yl)ethyl sulfide) and all other reagents were purchased from Sigma (St. Louis, MO)

Aboud O.A., Habib S.L., Trott J., Stewart B., Liang S., Chaudhari A.J., Sutcliffe J, & Weiss R.H. (2017). Glutamine addiction in kidney cancer suppresses oxidative stress and can be exploited for real-time imaging. Cancer research, 77(23), 6746-6758.

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Protein Expression Analysis in Lung Cancer

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Cells were lysed with RIPA buffer and proteins were separated on 13% SDS-PAGE and transferred onto a PVDF membrane. The membrane was stained immunochemically using anti-KEAP1, NRF2, NQO1, TXNRD1 (Proteintech, Chicago, IL, USA), and by using anti-AKR1C1 (Abnova, Taiwan). H720 and H727 cells were cultured in RPMI 1640, supplemented with 10% FBS and antibiotics. Equal amounts of protein lysates (35 μg) were resolved by SDS-PAGE (13% acrylamide) under reducing conditions and transferred onto PVDF membranes (Immobilon PVDF, Millipore, Billerica, MA, USA). After transfer, PVDF membranes were blocked with 5% non-fat milk and incubated with primary antibody overnight at 4 °C. Rabbit anti-NRF2 (Proteintech), rabbit anti-NQO1 (Proteintech), rabbit anti-TXNRD1 (Proteintech), mouse anti-AKR1C1 (Abnova) and anti-Actin (Sigma-Aldrich, St. Louis, MO, USA) were incubated overnight at 4 °C. PVDF membranes were incubated with ECL plus solution following the manufacturer’s instructions (Thermo Fisher Scientific, Rockford, IL, USA). Chemiluminescence signals were acquired by ChemiDoc XRS (Biorad, Hercules, CA, USA).

Sparaneo A., Fabrizio F.P., la Torre A., Graziano P., Di Maio M., Fontana A., Bisceglia M., Rossi A., Pizzolitto S., De Maglio G., Tancredi A., Grimaldi F., Balsamo T., Centra F., Manzorra M.C., Trombetta D., Pantalone A., Bonfitto A., Maiello E., Fazio V.M, & Muscarella L.A. (2019). Effects of KEAP1 Silencing on the Regulation of NRF2 Activity in Neuroendocrine Lung Tumors. International Journal of Molecular Sciences, 20(10), 2531.

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Quantitative α-SMA Protein Analysis

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Chemiluminescence western blot techniques were used to assess the α-SMA protein relative to the total protein (Ponceau stain). Polyvinylidene difluoride (PVDF) membranes (Bio-Rad) were used and probed for the primary α-SMA antibody (Abcam, 1:1000) diluted in blocking solution (5% skim milk) at 4 °C. Membranes were incubated with a secondary goat-anti-mouse horseradish peroxidase conjugated antibody (Jackson ImmunoResearch, 1:5000) for 1 h at RT. The chemiluminescence signal was detected using Pierce enhanced chemiluminescence (ECL) Plus solution (ThermoFisher Scientific) and an ImageQuant LAS 4000 detector.

Gonzalez Rodriguez A., Schroeder M.E., Walker C.J, & Anseth K.S. (2018). FGF-2 inhibits contractile properties of valvular interstitial cell myofibroblasts encapsulated in 3D MMP-degradable hydrogels. APL Bioengineering, 2(4), 046104.

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