Supersignal west pico dura chemiluminescent substrate

Manufactured by Thermo Fisher Scientific

Sourced in United States

SuperSignal West Pico/Dura Chemiluminescent Substrate is a laboratory reagent used for the detection and quantification of proteins in Western blot analysis. It utilizes a light-emitting chemical reaction to visualize the target proteins on a membrane.

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Lab products found in correlation

Blocking one p, by Nacalai Tesque (1 mentions) Phosphatase inhibitor cocktail, by Nacalai Tesque (1 mentions) Protease inhibitor cocktail, by Merck Group (1 mentions) Secondary antibodies conjugated to hrp, by Jackson ImmunoResearch (1 mentions) Pvdf membrane, by Merck Group (1 mentions) Cellytic m, by Merck Group (1 mentions) Fl 393, by Santa Cruz Biotechnology (1 mentions) Immobilon, by Merck Group (1 mentions) β actin, by Cell Signaling Technology (1 mentions)

Spelling variants of this product

SuperSignal West Pico (704 citations) Chemiluminescent substrate (358 citations) SuperSignal West Dura (232 citations) SuperSignal Chemiluminescent Substrate (135 citations) SuperSignal West Substrate (27 citations) Dura Chemiluminescent Substrate (12 citations) SuperSignal West Dura Chemiluminescent (5 citations) SuperSignal West Pico or Dura chemiluminescent substrates (4 citations) SuperSignal substrates (3 citations) SuperSignal West Pico or West Dura chemiluminescent substrates (2 citations)

4 protocols using supersignal west pico dura chemiluminescent substrate

Western Blot Analysis of Protein Expression

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The cells were washed in ice‐cold PBS and lysed using RIPA buffer supplemented with a protease inhibitor cocktail (Sigma‐Aldrich, St. Louis, MO) and phosphatase inhibitor cocktail (Nacalai Tesque). For Western blot analysis, 10 μg total protein were loaded and run on 7.5–15% precast gradient polyacrylamide gels. The gels were transferred to nitrocellulose membranes at 20 V overnight. Membranes were blocked in 5% milk or Blocking One‐P (Nacalai Tesque) for 20 min and incubated with the primary antibody at 4°C overnight. The primary antibodies used for Western blot analysis are listed in Supporting Information Table S1. After washing, the membranes were incubated for 1 hr at room temperature with the secondary antibody. Specific protein bands were detected using the SuperSignal West Pico/Dura Chemiluminescent Substrate (Thermo Fisher Scientific, Waltham, MA).

Kato M., Onoyama I., Yoshida S., Cui L., Kawamura K., Kodama K., Hori E., Matsumura Y., Yagi H., Asanoma K., Yahata H., Itakura A., Takeda S, & Kato K. (2020). Dual‐specificity phosphatase 6 plays a critical role in the maintenance of a cancer stem‐like cell phenotype in human endometrial cancer. International Journal of Cancer, 147(7), 1987-1999.

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Western Blot Analysis of Chromatin Proteins

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For Western analysis of chromatin proteins, an equal number of sperm and J1 mESCs were resuspended in 1x Laemmli sample buffer (5% 2-mercaptoethanol, 0.002% bromophenol blue, 10% Glycerol, 2% SDS, 62.5 mM Tris-HCl pH 6.8). Samples were boiled for 5 min and supernatant was loaded onto SDS-PAGE 4–15% gels at a ratio of 12:1 sperm:mESCs. Because sperm are haploid, sperm lanes contain 6-fold more genome equivalents than lanes with mESCs. Membranes were blocked in TBST (20 mM Tris, pH7.4, 150 mM NaCl, 0.05% Tween 20) with 5% nonfat milk powder and incubated overnight with the antibodies described above. Membranes were washed 3 times with TBST and incubated with secondary antibodies-conjugated to HRP (1:5000, Jackson ImmunoResearch Laboratories) for 1 hr. After three more washes, the presence of different proteins was detected using SuperSignal West Pico/Dura Chemiluminescent substrate (Thermo Scientific).

Jung Y.H., Kremsky I., Gold H.B., Rowley M.J., Punyawai K., Buonanotte A., Lyu X., Bixler B.J., Chan A.W, & Corces V.G. (2019). Maintenance of CTCF- and transcription factor-mediated interactions from the gametes to the early mouse embryo. Molecular cell, 75(1), 154-171.e5.

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Western Blot Analysis of Protein Samples

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Lysates boiled in Laemmli buffer were resolved by SDS-PAGE and transferred to PVDF membranes (Millipore) in Tris-glycine transfer buffer and 20% methanol for 2 hr at 100 V. After blocking with 5 % skim milk or BSA in 1× TBST buffer (20mM Tris, pH7.4, 150 mM NaCl, 0.05% Tween 20), membranes were incubated overnight with primary antibody (detailed in key resources table). Membranes were washed thrice with TBST and incubated with secondary antibody conjugated to HRP (1:3000, Novex, Invitrogen) in blocking buffer for 1 hr. Following three rounds of TBST washes, proteins were assayed using SuperSignal West Pico/Dura Chemiluminescent substrate (Thermofisher Scientific). Quantification was performed with “record measurements” in Adobe Photoshop.

Tan J., Tan Y.Y., Ngian Z.K., Chong S.Y., Rao V.K., Wang J.W., Zeng X, & Ong C.T. (2024). ApoE maintains neuronal integrity via microRNA and H3K27me3-mediated repression. iScience, 27(3), 109231.

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Western Blot Analysis of p53 Expression

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The cells were washed in ice‐cold PBS and lysed using CelLytic M (Sigma) following the manufacturer's instructions. Total protein (30 μg) was electrophoresed on 10% SDS‐polyacrylamide gels and transferred to nitrocellulose membranes (Immobilon; Merck Millipore, Darmstadt, Germany). The membranes were blocked and incubated overnight at 4°C with primary antibodies targeting p53 (FL‐393, 1:1000; Santa Cruz Biotechnology, Santa Cruz, CA, USA) and β‐actin (#4967, 1:1000; Cell Signaling, Danvers, MA, USA). After washing, membranes were incubated for 1 h with anti‐rabbit antibody (Sigma) diluted 1:4000. Specific protein bands were detected using the SuperSignal West Pico/Dura Chemiluminescent Substrate (Thermo Fisher Scientific, Waltham, MA, USA).

Kitade S., Onoyama I., Kobayashi H., Yagi H., Yoshida S., Kato M., Tsunematsu R., Asanoma K., Sonoda K., Wake N., Hata K., Nakayama K.I, & Kato K. (2016). FBXW7 is involved in the acquisition of the malignant phenotype in epithelial ovarian tumors. Cancer Science, 107(10), 1399-1405.

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