Eighty micrograms of total proteins from pool of three samples per each experimental group were subjected to 10% or 12.5% SDS-PAGE according to the method previously described [26 (link)]. Western blotting was performed using the primary polyclonal rabbit anti-mouse Sod1, Sod2, Cat, Gpx-1, Ho-1 or Nrf-2 antibody (Abcam, Cambridge, UK), following donkey anti-rabbit IgG horseradish peroxidase-conjugated secondary antibody (Amersham Biosciences Inc., USA). Anti-ERK-2 (C-14, Santa Cruz Biotechnology, Inc., USA) was used as a loading control. After the immunodetection the chemiluminescence signals were detected with the Alliance 4.7 Imaging System (UVITEC, Cambridge, UK).
Donkey anti rabbit igg horseradish peroxidase conjugated secondary antibody
Manufactured by Cytiva
Sourced in United Kingdom
Donkey anti-rabbit IgG horseradish peroxidase-conjugated secondary antibody is a laboratory reagent designed for detection and quantification of rabbit primary antibodies in various immunoassays. It consists of donkey-derived polyclonal antibodies that specifically bind to rabbit immunoglobulin G (IgG) molecules, conjugated with the enzyme horseradish peroxidase (HRP).
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2 protocols using donkey anti rabbit igg horseradish peroxidase conjugated secondary antibody
1
Oxidative Stress Protein Analysis
2
Western Blot Analysis of HO-1 and HO-2
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Eighty micrograms of post-mitochondrial supernatant from each sample (three pools, each of four mouse lungs) were subjected to 12.5% SDS-PAGE according to the method previously described [20] (link). Western blotting was performed using the primary polyclonal rabbit anti-mouse HO-1 or HO-2 antibody (Abcam, Cambridge,UK), following donkey anti-rabbit IgG, horseradish peroxidase-conjugated, secondary antibody (Amersham Biosciences Inc., USA). Anti-ERK-2 (C-14, Santa Cruz Biotechnology, Inc., USA), was used as a loading control [12] (link). The chemiluminescence signals were detected with the Alliance 4.7 Imaging System (UVITEC, Cambridge, UK).
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