ab181658 was purified from monospecific antiserum by a multi-step process which includes delipidation, salt fractionation and ion exchange chromatography followed by extensive dialysis against the buffer stated above.
Horseradish Peroxidase (HRP) is an enzyme commonly used as an indicator for chemical reactions which produce peroxide. The enzyme is routinely conjugated to antibodies for use in enzyme-based immunoassay systems.
HRP functions in the removal of H2O2 (hydrogen peroxide), oxidation of toxic reductants, biosynthesis and degradation of lignin, suberization, auxin catabolism, response to environmental stresses such as wounding, pathogen attack and oxidative stress. These functions might be dependent on each isozyme/isoform in each plant tissue.
Secreted Probable. Vacuole Probable. Note: Carboxy-terminal extension appears to target the protein to vacuoles.
All lanes : Anti-HRP antibody (ab181658) at 1/5000 dilution
Lane 1 : Horseradish peroxidase (reducing conditions) at 1 µg Lane 2 : Horseradish peroxidase (reducing conditions) at 0.25 µg Lane 3 : Horseradish peroxidase (non-reducing conditions) at 1 µg Lane 4 : Horseradish peroxidase (non-reducing conditions) at 0.25 µg
Secondary All lanes : Dylight 649 conjugated Donkey anti Goat at 1/10000 dilution
Predicted band size: 39 kDa
Reduced samples of purified Horseradish peroxidase contained 4% BME and were boiled for 5 minutes.
Western blot - Anti-HRP antibody (ab181658)
Anti-HRP antibody (ab181658) at 1/1000 dilution + Peroxidase at 0.05 µg
Secondary DyLight™ 649 goat secondary at 1/20000 dilution
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