Hydrostatin-SN1 was kindly provided by School of Pharmacy, Second Military Medical University, Shanghai, China. LPS was purchased from Sigma (St. Louis, MO, USA). Cell culture reagents were purchased from Invitrogen (Carlsbad, CA, USA). Enzyme-linked immunosorbent assay (ELISA) kits of TNF-α, IL-6, and IL-1β were obtained from R&D Systems (Minneapolis, MN, USA). The ECL Chemiluminescence kit and bicinchoninic acid (BCA) protein assay were purchased from Thermo Fisher Scientific Inc. (Waltham, MA, USA). The rabbit monoclonal antibodies for extracellular-signal related kinase 1/2 (ERK1/2), phospho-ERK1/2, NF-κBp65, phospho-NF-κBp65, IκBα and mouse monoclonal antibody for GAPDH were purchased from Cell Signaling Technology Inc. (Beverly, MA, USA). The horseradish peroxidase-conjugated goat anti-rabbit and goat anti-mouse secondary antibodies were provided by Santa Cruz Biotechnology (Dallas, TX, USA). Trizol reagent was purchased from Invitrogen (Carlsbad, CA, USA). Red blood cell lysis buffer and myeloperoxidase (MPO) assays were purchased from Beyotime Institute of Biotechnology (Jiangsu, China).
Horseradish peroxidase conjugated goat anti rabbit and goat anti mouse secondary antibodies
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Horseradish peroxidase-conjugated goat anti-rabbit and goat anti-mouse secondary antibodies are laboratory reagents used in various immunoassay techniques, such as Western blotting and immunohistochemistry. These antibodies are designed to bind and detect primary antibodies raised in rabbits or mice, and the horseradish peroxidase enzyme conjugated to them can be used to generate a signal for visualization and quantification.
Automatically generated - may contain errors
Lab products found in correlation
Cell culture reagents, by Thermo Fisher Scientific (1 mentions)
Bicinchoninic acid bca protein assay, by Thermo Fisher Scientific (1 mentions)
Ecl chemiluminescence kit, by Thermo Fisher Scientific (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Phospho nf κb p65, by Cell Signaling Technology (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
Red blood cell lysis buffer, by Beyotime (1 mentions)
Rabbit monoclonal anti p53, by Cell Signaling Technology (1 mentions)
Mouse monoclonal anti gapdh, by Cell Signaling Technology (1 mentions)
Rabbit monoclonal anti ago2, by Cell Signaling Technology (1 mentions)
Proteojet mammalian cell lysis reagent, by Thermo Fisher Scientific (1 mentions)
Supersignal detection reagents, by Thermo Fisher Scientific (1 mentions)
Anti β actin, by Merck Group (1 mentions)
Anti foxo3, by Cell Signaling Technology (1 mentions)
Anti flag hrp, by Abcam (1 mentions)
Anti survivin, by R&D Systems (1 mentions)
Anti dna pkcs, by Thermo Fisher Scientific (1 mentions)
Anti flag, by Cell Signaling Technology (1 mentions)
Dynabeads protein g, by Thermo Fisher Scientific (1 mentions)
Anti gfp, by Abcam (1 mentions)
3 protocols using horseradish peroxidase conjugated goat anti rabbit and goat anti mouse secondary antibodies
1
Anti-Inflammatory Effects of Hydrostatin-SN1
2
Western Blot Analysis of Cellular Proteins
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Whole-cell lysates were obtained using the ProteoJET Mammalian Cell Lysis Reagent (Thermo Scientific, Rockford, IL, USA). Proteins (50 μg) were separated on 10% SDS-PAGE gels and transferred to nitrocellulose membranes. The membranes were blocked with 5% non-fat dried milk in TBST (50 mM Tris (pH 7.5), 200 mM NaCl, 0.05% Tween 20) at room temperature for 1 h. The membranes were then incubated with the primary antibody in the blocking solution for 1 h at room temperature, washed three times with TBST for 15 min, incubated with the HRP-conjugated secondary antibody at room temperature for 1 h and then washed three times with TBST. Antigen-antibody complexes were detected using the SuperSignal detection reagents (Thermo Scientific, Rockford, IL, USA). The following antibodies were used: rabbit monoclonal anti-p53, rabbit monoclonal anti-AGO2, rabbit monoclonal anti-GADD45A, mouse monoclonal anti-GAPDH (Cell Signalling Technology, MA, USA) and horseradish peroxidase-conjugated goat-anti-rabbit and goat-anti-mouse secondary antibodies (Santa Cruz. CA, USA).
3
Ionizing Radiation Induced DNA-PKcs Signaling
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SW480 cells were transfected with survivin/PI3K expression vectors for 48 hours and cell lysates were prepared at 1 hour after a 4 Gy irradiation. Equal amounts of lysates were added to coupled Dynabeads Protein G (Thermo Fisher Scientific) and antibody complexes: Anti-GFP (#ab290; Abcam, RRID:AB_303395), anti-Flag (#2368S; Cell Signaling Technology), anti-DNA-PKcs (#MS-369-P1; Thermo Fisher Scientific) or IgG (Mouse #SC-2025, Rabbit #SC-2027; Santa Cruz Biotechnology) and incubated in a rotating mixer at 5 rpm overnight at 4 C. Next, beads were washed with PBS and proteins were eluted by boiling, subjected to Western immunoblotting and detected with primary antibodies: anti-survivin (AF886; R&D Systems), Anti-GFP (ab290; Abcam), anti-DNA-PKcs (#MS-369-P1; Thermo Fisher Scientific), anti-Flag-HRP (#ab49763; Abcam), anti-b-actin (A5441-.2ML; Sigma-Aldrich), anti-Foxo3 (#2497S), or anti-Foxo3 pS253 (#9466S; Cell Signaling Technology). For detection, horseradish peroxidase-conjugated goat anti-rabbit and goat anti-mouse secondary antibodies (Santa Cruz Biotechnology) and LI-COR WesternSure Premium Chemiluminescent Substrate (LI-COR) were used.
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