HC04 cells were either not treated or treated with 125 nM bafilomycin A1 for 2 h before mock infection or infection with B. pseudomallei at MOI 10. Cells and bacteria were co-cultured under standard conditions for the times indicated after which cells were washed with 1× PBS pH 7.4, and total proteins extracted. The expression levels of LC3-II and Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH) were determined by western blotting using a rabbit anti-LC3 polyclonal antibody (2775S; Cell Signaling Technology) and a mouse anti-GAPDH monoclonal antibody (611463; BD Pharmacia, San Jose, CA, USA) followed by a Horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG antibody (sc2004; Santa Cruz Biotechnology, Inc.) and a HRP-conjugated goat anti-mouse IgG antibody (sc2005; Santa Cruz Biotechnology, Inc.). Signal was observed using Luminata Forte Western HRP substrate (WBLUF0100; Merck Millipore, Darmstadt, Germany). Band intensities were quantitated using the ImageJ software.
Hrp conjugated goat anti mouse igg antibody sc2005
Manufactured by Santa Cruz Biotechnology
Sourced in United States
The HRP-conjugated goat anti-mouse IgG antibody (sc-2005) is a secondary antibody conjugated with horseradish peroxidase (HRP). It is designed to detect and bind to mouse immunoglobulin G (IgG) antibodies in various immunoassays and experimental applications.
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2 protocols using hrp conjugated goat anti mouse igg antibody sc2005
1
Bafilomycin A1 Modulates Autophagy in B. pseudomallei Infection
2
Quantification of Autophagy Markers
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U937 cells were co-cultured with bacteria under standard conditions following specified time point, then cells were washed with 1X PBS pH 7.4, and total proteins were extracted using RIPA buffer (Thermo Fisher Scientific, MA, USA). The expression levels of LC3-II and Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH) were determined by western blotting using a rabbit anti-LC3 polyclonal antibody (2775S; Cell Signaling Technology, MA, USA) and a mouse anti-GAPDH monoclonal antibody (611,463; BD Pharmacia, San Jose, CA, USA), and followed by a Horseradish peroxidase (HRP)-conjugated goat anti- rabbit IgG antibody (sc2004; Santa Cruz Biotechnology, Inc., TX, USA) and a HRP-conjugated goat anti-mouse IgG antibody (sc2005; Santa Cruz Biotechnology, Inc., TX, USA). The signal was visualized using Pierce® ECL Western blotting substrate (Thermo Fisher Scientific, MA, USA).
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