Mast cells were lysed (ice/30 min) in 350 µL of lysis buffer (50 mM Hepes pH 7.4, 250 mM NaCl, 20 mM NaF, 10 mM iodoacetamide, 0.5% (w/v) Triton X100, 1 mM PMSF (phenylmethylsulfonylfluoride), 500 µg/ml aprotinin, 1.0 mg/ml leupeptin and 2.0 mg/ml chymostatin). Lysates were clarified (17,000 g, 20 min) and acetone precipitated (1.4 volumes acetone, 1 h/−20°C, followed by 10,000 g, 5 min). Protein was resolved by 10% reducing SDS-PAGE in a modified Laemmli buffer and electro-transferred to PVDF in 192 mM glycine, 25 mM Tris (pH 8.8). Membranes were blocked (5% non-fat milk in PBS, 1 h, RT) and probed (primary antibodies in PBS/0.05% Tween-20/0.05% NaN3, 16 h/4°C). Developing antibodies comprised anti-rabbit or anti-mouse IgGs conjugated to horseradish peroxidase (Amersham) at 0.1 µg/ml in PBS/0.05% Tween-20 (45 min/RT). Signal was visualized using ECL (Amersham) and Kodak BioMax film. Films were scanned at >600 dpi and quantification was performed using Image J (NIH).
Anti rabbit or anti mouse iggs conjugated to horseradish peroxidase
Manufactured by Cytiva
Anti-rabbit or anti-mouse IgGs conjugated to horseradish peroxidase are lab equipment used for detection and quantification in immunoassays. They serve as secondary antibodies that bind to primary antibodies targeting rabbit or mouse antigens, and the horseradish peroxidase enzyme allows for colorimetric or chemiluminescent signal generation.
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Lab products found in correlation
2 protocols using anti rabbit or anti mouse iggs conjugated to horseradish peroxidase
1
Mast Cell Protein Extraction and Analysis
2
Mast Cell Protein Extraction and Analysis
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Mast cells were lysed (ice/30 min) in 350 µL of lysis buffer (50 mM Hepes pH 7.4, 250 mM NaCl, 20 mM NaF, 10 mM iodoacetamide, 0.5% (w/v) Triton X100, 1 mM PMSF (phenylmethylsulfonylfluoride), 500 µg/ml aprotinin, 1.0 mg/ml leupeptin and 2.0 mg/ml chymostatin). Lysates were clarified (17,000 g, 20 min) and acetone precipitated (1.4 volumes acetone, 1 h/−20°C, followed by 10,000 g, 5 min). Protein was resolved by 10% reducing SDS-PAGE in a modified Laemmli buffer and electro-transferred to PVDF in 192 mM glycine, 25 mM Tris (pH 8.8). Membranes were blocked (5% non-fat milk in PBS, 1 h, RT) and probed (primary antibodies in PBS/0.05% Tween-20/0.05% NaN3, 16 h/4°C). Developing antibodies comprised anti-rabbit or anti-mouse IgGs conjugated to horseradish peroxidase (Amersham) at 0.1 µg/ml in PBS/0.05% Tween-20 (45 min/RT). Signal was visualized using ECL (Amersham) and Kodak BioMax film. Films were scanned at >600 dpi and quantification was performed using Image J (NIH).
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