The mAb used include: C6G9 (Sigma-Aldrich, St. Louis, MO, USA) anti-human CEA (CD66e) and Tetra-His (Qiagen, GmbH, Hilden, Germany). The polyclonal antibodies included: phycoerytrin (PE)-conjugated goat F(ab’)2 fragment anti-mouse IgG (Fc fragment specific, Jackson Immuno Research, Newmarket, UK), horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG (Fc specific) (Sigma-Aldrich), and IRDye800-conjugated donkey anti-mouse IgG (H&L) (Rockland Immunochemicals, Gilbertsville, PA, USA). Human CEA was obtained from Calbiochem (Merck, Darmstadt, Germany) and bovine serum albumin (BSA) was from Sigma-Aldrich.
Horseradish peroxidase hrp conjugated goat anti mouse igg fc specific
Manufactured by Merck Group
Sourced in United Kingdom, Germany
Horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG (Fc specific) is a laboratory reagent used to detect and quantify mouse immunoglobulin G (IgG) in various immunoassays. The HRP enzyme is covalently attached to the goat-derived antibody, which specifically binds to the Fc region of mouse IgG. This conjugate can be used as a secondary antibody in techniques such as enzyme-linked immunosorbent assay (ELISA), western blotting, and immunohistochemistry to amplify the signal and facilitate the visualization of target mouse IgG proteins.
Automatically generated - may contain errors
Lab products found in correlation
Irdye800 conjugated donkey anti mouse igg h l, by Rockland Immunochemicals (1 mentions)
Tetra his, by Qiagen (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Anti his mab, by Qiagen (1 mentions)
Chemidoc mp imaging system machine, by Bio-Rad (1 mentions)
Tris glycine gel, by Thermo Fisher Scientific (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Pierce ecl western blotting substrate, by Bio-Rad (1 mentions)
3 protocols using horseradish peroxidase hrp conjugated goat anti mouse igg fc specific
1
Antibody Characterization for CEA Detection
2
Western Blot Protein Detection
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Samples were separated under reducing conditions on 10%–20% Tris-glycine gels (Life Technologies, Carlsbad, California, USA), transferred onto PVDF membranes (Merck Millipore, Tullagreen, Carrigtwohill, Ireland) and probed with 200 ng/mL anti-His mAb (Qiagen, Hilden Germany), followed by incubation with 1.6 µg/mL horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG, Fc specific (Sigma-Aldrich). Visualization of protein bands was performed with Pierce ECL Western Blotting substrate (Rockford, IL, USA) and ChemiDoc MP Imaging System machine (Bio-Rad Laboratories, Hercules, California, USA).
3
CEA Binding Assay with MFE23 Variants
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The ability of unlabeled or 124 I-labelled MFE23-scFv and MFE23 N -trimerbody to bind CEA was analyzed by ELISA as previously described, with minor modifications. 23 In brief, Maxisorp (NUNC Brand Products, Roskilde, Denmark) plates were coated (0.3 µg/well) with human CEA (Sigma-Aldrich) and after washing and blocking with 200 μl 5% BSA in PBS, 100 μl with indicated amount of protein was added for 1 hour at room temperature. After three washes, 100 μl of anti-myc 9E10 mAb (1 μg/ml) were added for 1 hour at room temperature. After removal of detection antibody, 100 μl of horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG (Fc specific; Sigma-Aldrich) were added for 1 hour at room temperature, after which the plate was washed and developed. Antigen titration was performed by triplicate with serial dilutions of both antibodies (concentration ranges 0.02-100 nM). Binding constants EC 50 and Kd were estimated from fitting curves to data using nonlinear regression according to the Graphpad Curve Fitting Guide (Graphpad Prism, version 6.01).
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