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Enhanced chemiluminescence ecl western blotting detection reagent

Manufactured by GE Healthcare
Sourced in United Kingdom

Enhanced chemiluminescence (ECL) Western blotting detection reagent is a laboratory product used to visualize and detect proteins in Western blot analysis. It provides a sensitive and reliable method for the detection of target proteins by generating a chemiluminescent signal that can be captured and analyzed.

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4 protocols using enhanced chemiluminescence ecl western blotting detection reagent

1

Cellular Response to Inflammatory Stimuli

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Dulbecco’s modified Eagle medium (DMEM), fetal bovine serum (FBS), L-glutamine/penicillin/streptomycin, and anti-α-tubulin monoclonal antibodies (mAbs) were purchased from Invitrogen (Thermo Fisher Scientific, Waltham, MA, USA). LPS and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were bought from Sigma-Aldrich (St. Louis, MO, USA). Anti-iNOS, COX2, and Nrf2 polyclonal antibodies (pAbs) were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). An anti-HO-1 pAb was purchased from Enzo (Farmingdale, New York, NY, USA). The anti-phospho-p65 (Ser536) mAb was purchased from Cell Signaling (Danvers, MA, USA). Horseradish peroxidase (HRP)-conjugated donkey anti-rabbit immunoglobulin G (IgG) and sheep anti-mouse IgG were purchased from Amersham (Buckinghamshire, U.K.). Enhanced chemiluminescence (ECL) Western blotting detection reagent and Hybond-P polyvinylidene difluoride (PVDF) blotting membranes were purchased from GE Healthcare Life Sciences (Waukesha, WI, USA).
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2

Collagen and Thrombogenic Pathway Activation

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Collagen (type I), 9, 11-dideoxy-11α, 9α-epoxymethanoprostaglandin (U46619), and thrombin were purchased from Chrono-Log Corporation (Havertown, PA, USA). Anti-phospho-p38 mitogen-activated protein kinase (MAPK) (Thr180/Tyr182), anti-phospho-c-Jun N-terminal kinase (JNK) (Thr183/Tyr185), anti-phospho-(Ser) protein kinase C (PKC) substrate, anti-JNK polyclonal antibodies (pAb), and anti-p38 MAPK and anti-Akt monoclonal antibodies (mAb) were purchased from Cell Signaling Technology (Beverly, MA, USA). Anti-phospho-Akt (Ser473) pAb was purchased from Biorbyt (Cambridge, UK), and anti-pleckstrin (p47) pAb was purchased from Gene Tex (Irvine, CA, USA). Hybond-P polyvinylidene difluoride (PVDF) membranes, enhanced chemiluminescence (ECL) Western blotting detection reagent, and the analysis system were purchased from GE Healthcare Life Sciences (Buckinghamshire, UK). Horseradish peroxidase-conjugated goat anti-rabbit and anti-mouse immunoglobulin G antibodies were obtained from Jackson ImmunoResearch Laboratories (West Grove, PA, USA).
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3

Compound Interactions with Serum Proteins

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Catechins, DHA, p-aminobenzoic acid (ABA), and human serum albumin (HSA) (fatty acid and globulin free, purity 99%) were obtained from Sigma-Aldrich. O-Methyl derivatives of EGCG were prepared as previously reported [14 (link)]. Bovine serum albumin (BSA) and pyrroloquinoline quinone (PQQ) were obtained from Wako Pure Chemicals (Japan). The HRP-linked anti-mouse IgM (for ELISA analysis) was obtained from Cappel Laboratories. Hydrogen peroxide (31%, W/V) was obtained from Mitsubishi Gas Co., Ltd. Horseradish peroxidase (HRP)-conjugated NeutrAvidin and enhanced chemiluminescence (ECL) Western blotting detection reagents were obtained from GE Healthcare. All of the other reagents used in the study were of analytical grade and obtained from commercial sources.
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4

Colorectal Carcinoma Cell Line RKO Assay

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Human colorectal carcinoma cell line, RKO, was obtained from American Type Culture Collection (Manassas, Virginia, USA). Minimum essential medium (MEM) was purchased from Gibco Laboratories (Paisley, UK). Fetal bovine serum (FBS) and [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] (MTT) were purchased from Sigma-Aldrich Chemical Company (Dorset, UK). Six-well flat-bottom tissue culture plates were purchased from Nunc Inc. (Hereford, UK). Phenylmethylsulfonyl fluoride (PMSF) was purchased from Sigma-Aldrich Co., Ltd. (St. Louis, MO, USA) and protease inhibitor mixture solution was obtained from Wako Pure Chemical Industries, Ltd. (Osaka, Japan). Enhanced chemiluminescence (ECL) western blotting detection reagents and pure nitrocellulose MEMbranes were purchased from GE Healthcare (Buckinghamshire, UK). Primary antibodies for the detection of caspase 3 and β-actin and goat antirabbit and goat anti-mouse IgG horseradish peroxidase (HRP)-conjugated secondary antibodies were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). All chemicals and reagents were of analytical grade and purchased from Sigma-Aldrich Co., Ltd. unless otherwise specified.
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