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Supersignal west pico and femto chemiluminescent substrates

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SuperSignal West Pico and Femto chemiluminescent substrates are products designed for the detection of proteins in Western blotting applications. They provide a sensitive and reliable method for visualizing target proteins through the generation of a chemiluminescent signal.

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Lab products found in correlation

Bca protein assay kit, by Thermo Fisher Scientific (6 mentions) Protease and phosphatase inhibitor tablet, by Roche (3 mentions) Ripa lysis and extraction buffer, by Thermo Fisher Scientific (3 mentions) Superscript 2 reverse transcriptase, by Thermo Fisher Scientific (2 mentions) Anti snat1 slc38a1 d9l2p, by Cell Signaling Technology (2 mentions) Anti pgc 1α h 300 and anti snat2 c 6, by Santa Cruz Biotechnology (2 mentions) Mg132, by Merck Group (2 mentions) Ne per nuclear and cytoplasmic extraction reagent, by Thermo Fisher Scientific (2 mentions) Dibutyryl camp db camp, by Merck Group (1 mentions) Lab tek chambered coverglass, by Thermo Fisher Scientific (1 mentions) Rolipram, by Cayman Chemical (1 mentions) Forskolin, by Merck Group (1 mentions) Ce3f4, by Bio-Techne (1 mentions) Rp camp, by Merck Group (1 mentions) Mem glutamax, by Thermo Fisher Scientific (1 mentions) Ecl prime western blotting detection reagent, by GE Healthcare (1 mentions) Amersham hybond p, by GE Healthcare (1 mentions) Anti tbk1, by Cell Signaling Technology (1 mentions) Anti p tbk1, by Cell Signaling Technology (1 mentions) Anti β actin, by Cell Signaling Technology (1 mentions)

Close spelling variants of this product

SuperSignal West Pico Chemiluminescent Substrate (2569 citations) SuperSignal West Pico (704 citations) SuperSignal West Femto (427 citations) Chemiluminescent substrate (358 citations) SuperSignal West Femto Chemiluminescent Substrate (315 citations) SuperSignal West Femto Substrate (258 citations) SuperSignal West Pico Substrate (146 citations) SuperSignal Chemiluminescent Substrate (135 citations) Femto Chemiluminescent Substrate (89 citations) West Pico Chemiluminescent Substrate (56 citations)

6 protocols using supersignal west pico and femto chemiluminescent substrates

1

Reagents and Materials for Cell Culture Protocols

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Chemicals and other materials were obtained from the following sources: hydrogen peroxide (H2O2), dibutyryl-cAMP (dbcAMP), and H89 from Sigma, St. Louis, MO, USA; 8CPT-6phenyl-cAMP from Biolog; DMEM/F12 (1 : 1), MEM+GlutaMax™, 100x penicillin/streptomycin (Pen/Strep), and fetal bovine serum from Thermo Fisher Scientific, Waltham, MA, USA; Superscript™ II reverse transcriptase from Invitrogen; RNase inhibitor, dNTP mixture, and oligo (dT) from Promega; polymerase chain reaction (PCR) master mix, protein size marker, and SuperSignal® West Pico and Femto chemiluminescent substrates from Thermo Fisher Scientific; and Amersham Hybond™-P polyvinylidenedifluoride (PVDF) membrane and ECL™ Prime Western Blotting Detection Reagent from GE Healthcare (Chicago, IL, USA).
Ju W.K., Shim M.S., Kim K.Y., Park T.L., Ahn S., Edwards G, & Weinreb R.N. (2019). Inhibition of cAMP/PKA Pathway Protects Optic Nerve Head Astrocytes against Oxidative Stress by Akt/Bax Phosphorylation-Mediated Mfn1/2 Oligomerization. Oxidative Medicine and Cellular Longevity, 2019, 8060962.
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2

Investigating IGF-1 Signaling Pathways

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Chemicals and other materials were obtained from the following sources: H2O2, forskolin, Rp-cAMP, and H89 from Sigma; Rolipram from Cayman Chemical; Recombinant Rat IGF-1 from Prospec; DMEM/F12 (1:1), MEM+GlutaMax, 100 × penicillin–streptomycin (Pen/Strep) and fetal bovine serum from Gibco; Superscript II reverse transcriptase from Invitrogen; RNase inhibitor, dNTP mixture and oligo (dT) from Promega; PCR master mix, protein size marker, SuperSignal West Pico and Femto chemiluminescent substrates from Thermo scientific; Amersham Hybond-P (PVDF membrane) and ECL Prime Western Blotting Detection Reagent from GE Healthcare; Lab-Tek Chambered cover glass from Nunc; CE3F4 from TOCRIS.
Shim M.S., Kim K.Y., Bu J.H., Nam H.S., Jeong S.W., Park T.L., Ellisman M.H., Weinreb R.N, & Ju W.K. (2018). Elevated intracellular cAMP exacerbates vulnerability to oxidative stress in optic nerve head astrocytes. Cell Death & Disease, 9(3), 285.
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3

Western Blot Analysis of T Cell Proteins

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T cells were washed twice in 1X cold PBS and cell pellets were lysed using RIPA lysis and extraction buffer (Thermo Fisher Scientific) supplemented with a protease and phosphatase inhibitor tablet (Roche). Homogenates were centrifuged at 14,000 rpm for 30 min at 4°C, and the supernatants were collected. Protein concentrations were determined using a BCA protein assay kit (Thermo Fisher Scientific). Equivalent amounts of protein were separated via SDS-PAGE and transferred onto PVDF membranes following the standard protocol. Following antibodies were used: anti-GLUT1 (D3J3A), anti-β-ACTIN (4967S), anti-ASCT2 (V501), and anti-SNAT1/SLC38A1 (D9L2P) (Cell Signaling Technologies); anti-PGC-1α (H-300) and anti-SNAT2 (C-6) (Santa Cruz Biotechnology); and goat anti-rabbit and mouse secondary antibodies conjugated with HRP (Thermo Fisher Scientific). SuperSignal West Pico and Femto chemiluminescent substrates (Thermo Fisher Scientific) were used to image blots in a FluorChemE instrument (ProteinSimple). The proteasomal inhibitor MG132 (10 μM, Sigma) and translation inhibitor cycloheximide (CHX, 50 μg/ml, Sigma) were used to assess protein abundance and stability. Densitometric quantification was performed using Image J software (NIH).
Song M., Sandoval T.A., Chae C.S., Chopra S., Tan C., Rutkowski M.R., Raundhal M., Chaurio R.A., Payne K.K., Konrad C., Bettigole S.E., Shin H.R., Crowley M.J., Cerliani J.P., Kossenkov A.V., Motorykin I., Zhang S., Manfredi G., Zamarin D., Holcomb K., Rodriguez P.C., Rabinovich G.A., Conejo-Garcia J.R., Glimcher L.H, & Cubillos-Ruiz J.R. (2018). IRE1α-XBP1 controls T cell function in ovarian cancer by regulating mitochondrial activity. Nature, 562(7727), 423-428.
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4

Western Blot Analysis of T Cell Proteins

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T cells were washed twice in 1X cold PBS and cell pellets were lysed using RIPA lysis and extraction buffer (Thermo Fisher Scientific) supplemented with a protease and phosphatase inhibitor tablet (Roche). Homogenates were centrifuged at 14,000 rpm for 30 min at 4°C, and the supernatants were collected. Protein concentrations were determined using a BCA protein assay kit (Thermo Fisher Scientific). Equivalent amounts of protein were separated via SDS-PAGE and transferred onto PVDF membranes following the standard protocol. Following antibodies were used: anti-GLUT1 (D3J3A), anti-β-ACTIN (4967S), anti-ASCT2 (V501), and anti-SNAT1/SLC38A1 (D9L2P) (Cell Signaling Technologies); anti-PGC-1α (H-300) and anti-SNAT2 (C-6) (Santa Cruz Biotechnology); and goat anti-rabbit and mouse secondary antibodies conjugated with HRP (Thermo Fisher Scientific). SuperSignal West Pico and Femto chemiluminescent substrates (Thermo Fisher Scientific) were used to image blots in a FluorChemE instrument (ProteinSimple). The proteasomal inhibitor MG132 (10 μM, Sigma) and translation inhibitor cycloheximide (CHX, 50 μg/ml, Sigma) were used to assess protein abundance and stability. Densitometric quantification was performed using Image J software (NIH).
Song M., Sandoval T.A., Chae C.S., Chopra S., Tan C., Rutkowski M.R., Raundhal M., Chaurio R.A., Payne K.K., Konrad C., Bettigole S.E., Shin H.R., Crowley M.J., Cerliani J.P., Kossenkov A.V., Motorykin I., Zhang S., Manfredi G., Zamarin D., Holcomb K., Rodriguez P.C., Rabinovich G.A., Conejo-Garcia J.R., Glimcher L.H, & Cubillos-Ruiz J.R. (2018). IRE1α-XBP1 controls T cell function in ovarian cancer by regulating mitochondrial activity. Nature, 562(7727), 423-428.
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5

Quantifying Protein Phosphorylation in BMDCs

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BMDCs were washed twice in 1X cold PBS and cell pellets were lysed using RIPA lysis and extraction buffer (Thermo Fisher Scientific) supplemented with a protease and phosphatase inhibitor tablet (Roche). Homogenates were centrifuged at 14,000 rpm for 30 min at 4°C, and the supernatants were collected. Protein concentrations were determined using a BCA protein assay kit (Thermo Fisher Scientific). Equivalent amounts of protein were separated via SDS-PAGE and transferred onto PVDF membranes following standard protocols. The following antibodies were used: anti-beta actin (Cell Signaling Technologies Cat# 4967), anti-pTBK1 (Cell Signaling Technologies Cat# 5438), anti-TBK1 (Cell Signaling Technologies Cat# 3504), anti-pIRE3 (Cell Signaling Technologies Cat# 29047), anti-IRE3 (Cell Signaling Technologies Cat# 4302), and goat anti-rabbit secondary antibodies conjugated with HRP (Thermo Fisher Scientific Cat# G-21234). SuperSignal West Pico and Femto chemiluminescent substrates (Thermo Fisher Scientific) were used to image blots in a FluorChemE instrument (ProteinSimple).
Chae C.S., Sandoval T.A., Hwang S.M., Park E.S., Giovanelli P., Awasthi D., Salvagno C., Emmanuelli A., Tan C., Chaudhary V., Casado J., Kossenkov A.V., Song M., Barrat F.J., Holcomb K., Romero-Sandoval E.A., Zamarin D., Pépin D., D’Andrea A.D., Färkkilä A, & Cubillos-Ruiz J.R. (2022). Tumor-derived Lysophosphatidic Acid Blunts Protective Type-I Interferon Responses in Ovarian Cancer. Cancer discovery, 12(8), 1904-1921.
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6

Detailed Reagents and Materials Protocol

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The following items were purchased: fetal bovine serum (FBS), DMEM, RPMI, MEM and trypsin solution from Gibco (Life Technologies); RNeasy Mini kit from Qiagen (Hilden, Germany); High Capacity RNA-to-cDNA kit from Applied Biosystems (Darmstadt, Germany); NE-PER Nuclear and Cytoplasmic Extraction reagents and SuperSignal West Pico and Femto chemiluminescent substrates from Thermo Fisher Scientific (Rockford, IL, USA); FuGENE HD Transfection reagent from Promega (Madison, WI, USA); G418 (Geneticin) from Roche Diagnostics Deutschland GmbH (Mannheim, Germany); laminin, collagen from human placenta type IV [(collagen IV) from Sigma-Aldrich]; Cell Counting kit-8 from Dojindo Laboratories (Kumamoto, Japan); BioCoat Matrigel Invasion Chamber (8-µm pore size) and fibronectin from BD Biosciences (Franklin Lakes, NJ, USA); gemcitabine hydrochloride from Wako Pure Chemical Industries (Osaka, Japan); Fluorescent Mounting Medium from Dako Japan (Tokyo, Japan); Histofine Simple Stain Max PO (M) or (R) kit from Nichirei Biosciences, Inc., (Tokyo, Japan); and recombinant human epidermal growth factor (EGF, 236-EG) and recombinant human platelet-derived growth factor (PDGF, 220-BB) from R&D Systems (Minneapolis, MN, USA).
SUMIYOSHI H., MATSUSHITA A., NAKAMURA Y., MATSUDA Y., ISHIWATA T., NAITO Z, & UCHIDA E. (2016). Suppression of STAT5b in pancreatic cancer cells leads to attenuated gemcitabine chemoresistance, adhesion and invasion. Oncology Reports, 35(6), 3216-3226.
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