Western blot analysis for human iPSC–derived cardiomyocytes was performed, as described previously (25 (link)), using antibodies to dystrophin (ab15277, Abcam; D8168, Sigma-Aldrich), glyceraldehyde-3-phosphate dehydrogenase (MAB374, Millipore), and cardiac myosin heavy chain (ab50967, Abcam). Goat anti-mouse and goat anti-rabbit horseradish peroxidase–conjugated secondary antibodies (Bio-Rad) were used for described experiments.
Goat anti mouse and goat anti rabbit horseradish peroxidase conjugated secondary antibodies
Manufactured by Bio-Rad
Sourced in United States, Canada
Goat anti-mouse and goat anti-rabbit horseradish peroxidase–conjugated secondary antibodies are detection reagents used in immunoassays and immunohistochemistry techniques. They bind to primary antibodies raised in mouse or rabbit and are conjugated with the enzyme horseradish peroxidase, enabling signal amplification and visualization.
Automatically generated - may contain errors
Lab products found in correlation
Ab15277, by Abcam (1 mentions)
Ab50967, by Abcam (1 mentions)
D8168, by Merck Group (1 mentions)
Mab374, by Merck Group (1 mentions)
Monoclonal anti ezrin, by BD (1 mentions)
Human tgf β1, by Merck Group (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Gsk650394, by Bio-Techne (1 mentions)
Mouse anti egfr, by Santa Cruz Biotechnology (1 mentions)
Mouse anti ezrin, by BD (1 mentions)
Phospho egfr y1068, by Cell Signaling Technology (1 mentions)
Phospho akt s473, by Cell Signaling Technology (1 mentions)
P44 42 mapk erk1 2, by Cell Signaling Technology (1 mentions)
Phospho erk1 2t202 y204, by Cell Signaling Technology (1 mentions)
Actin ac15, by Merck Group (1 mentions)
Phospho egfr, by Merck Group (1 mentions)
Ham s f 12k kaighn s medium, by Thermo Fisher Scientific (1 mentions)
Cho cells, by American Type Culture Collection (1 mentions)
4 protocols using goat anti mouse and goat anti rabbit horseradish peroxidase conjugated secondary antibodies
1
Analyzing iPSC-Derived Cardiomyocytes by Western Blot
2
Antibody-Based Analysis of Membrane Protein Localization
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The antibodies used were polyclonal anti-LMTK2 (SAB4500900, Sigma-Aldrich, St. Louis, MO, United States) and monoclonal anti-Na+/K+ ATPase a-1 (clone C464.6, Merck KGaA, Darmstadt, Germany), monoclonal anti-Rab5 (Cat. 610282) and monoclonal anti-Ezrin (Cat. 610603, BD Biosciences, San Jose, CA, United States), polyclonal anti-Rab11A (Cat. 71-5300, Thermo Fisher Scientific, Waltham, MA, United States), and goat anti-mouse and goat anti-rabbit horseradish peroxidase-conjugated secondary antibodies (Bio-Rad Laboratories, Hercules, CA, United States). All antibodies were used at the concentrations recommended by the manufacturer or as indicated in the figure captions. Human TGF-β1 (Sigma-Aldrich) was resuspended in the vehicle containing 4 mM HCl and 1 mg/mL bovine serum albumin (BSA, Sigma-Aldrich) and used at a concentration 15 ng/ml.
3
Antibodies and Reagents for CFTR Localization
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The antibodies used were: rabbit anti-SGK1 antibody (Sigma); mouse anti-CFTR C-terminus antibody (clone 24-1; R&D systems, Minneapolis, MN); mouse anti-CFTR antibody (clone 596; University of North Carolina Cystic Fibrosis Center, Chapel Hill, NC); mouse anti-ezrin, mouse anti-Rab5a and mouse anti-EEA1 antibodies (BD Biosciences, San Jose, CA); rabbit anti-Rab 11a (Life Technologies Corp); mouse anti-EGFR (Santa Cruz Biotechnology, Inc, Santa Cruz, CA); horseradish peroxidase-conjugated goat anti-mouse and goat anti-rabbit secondary antibodies (Bio-Rad, Hercules, CA). SGK1-S442D, a constitutively active form of human SGK1 and human SGK1-K127N, an inactive form of human SGK1, were constructed as described [15] . The SGK inhibitor GSK 650394 was obtained from Tocris (purchased through R&D Systems, Minneapolis, MN). All antibodies and reagents were used at the concentrations recommended by the manufacturers.
4
EGFR Signaling in CHO Cells
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CHO cells were purchased from American Type Culture Collection (ATCC) and maintained in 10% CO2 with Ham’s F-12K (Kaighn’s) medium (Gibco) supplemented with 10% FBS and 1% penicillin-streptomycin (both from Genessee Scientific). Antibodies recognizing the following proteins were purchased: Actin AC-15 (Sigma-Aldrich), EGFR, phospho-EGFR, phospho-EGFR Y1068, phospho-EGFR Y992, phospho-EGFR Y1045, Akt, phospho-Akt S473, p44/42 MAPK (Erk1/2), and phospho-Erk1/2 T202/Y204 (Cell Signaling Technologies). Horseradish peroxidase-conjugated goat anti-mouse and goat anti-rabbit secondary antibodies were purchased from Bio-Rad Laboratories. The primary antibodies, secondary antibodies, and dilutions are listed in Supplementary Table 16 .
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