Protein extraction and western blot were carried out as previously described (Ponz-Sarvise et al, 2011 (link)). The primary antibodies (diluted at 1 : 1000) were as follows: anti-human ITGα5 (Sigma), anti phospho-Src (Cell Signaling, Danvers, MA, USA) and anti-Src (Cell Signaling). An anti-human β-actin antibody (Sigma) at 1 : 10 000 dilution was used as a loading control. Horseradish peroxidase-labelled secondary antibodies (GE Healthcare, Waukesha, WI, USA) against the corresponding primary antibodies were used. Immunoreactive bands were visualised by a chemiluminescent method using the Lumi-lightPLUS kit (Roche, Palo Alto, CA, USA).
Horseradish peroxidase labelled secondary antibodies
Manufactured by GE Healthcare
Sourced in United States
Horseradish peroxidase-labelled secondary antibodies are laboratory reagents used in various immunoassay techniques. They consist of a secondary antibody that is conjugated with the enzyme horseradish peroxidase. These antibodies can be used to detect and quantify target proteins or other molecules in biological samples.
Automatically generated - may contain errors
2 protocols using horseradish peroxidase labelled secondary antibodies
1
Integrin α5 Protein Extraction and Western Blot
2
SDS-PAGE and Immunoblotting Protocols for Protein Analysis
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SDS‐PAGE and immunoblotting were performed following standard protocols and primary antibodies as follows. Secretion assays: α‐AvrBs3 (Knoop et al., 1991 ), α‐HrpF (Büttner et al., 2002 ), α‐HrcJ (Rossier et al., 2000 ). Translocation assays: α‐FLAG (mouse; Sigma‐Aldrich). Liposome flotation: α‐GST (goat; GE Healthcare). Fractionation: α‐GFP (mouse; Roche Diagnostics, Mannheim, Germany), α‐XopB (Schulze et al., 2012 ), α‐AcnB (Gruer et al., 1997 ), α‐HrcJ (Rossier et al., 2000 ). Horseradish peroxidase‐labelled secondary antibodies (GE Healthcare) were detected by enhanced chemiluminescence.
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