Anti-ABCA1 antibody [AB.H10] (ab18180)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-These buffers may be stored at 4°C for several weeks or aliquoted and stored at -20°C for up to a year.	-Mouse (1:500) -To reduce and denature your samples, boil each cell lysate in sample buffer at 100°C for 5 min. Lysates can be aliquoted and stored at -20°C for future use.

Upstream tips
-These buffers may be stored at 4°C for several weeks or aliquoted and stored at -20°C for up to a year.
Protocol tips
-Mouse (1:500) -To reduce and denature your samples, boil each cell lysate in sample buffer at 100°C for 5 min. Lysates can be aliquoted and stored at -20°C for future use.

Publication protocol

Liver lysates were prepared as previously described (Nagajyothi et al. 2014). An aliquot of each sample (40 µg protein) was subjected to a 4–15% gradient SDS-PAGE (except 14% gel used for LC3) and the proteins transferred to nitrocellulose filters for immunoblot analysis. p-AMPK beta specific rabbit monoclonal antibody (1:1000 dilution, Ser108, Cell Signaling), AMPK beta specific rabbit monoclonal antibody (1:1000 dilution, Cell Signaling), ABCA1 specific mouse monoclonal antibody (1:500 dilution, AB.H10, Abcam), p-mTOR specific rabbit (1:1000 dilution, Ser2448, Cell Signaling), LC3 B1/2 specific mouse (1:1000 dilution, M 186-3, MBL), p-62 specific rabbit (1:1000 dilution, PM045, MBL), Fatty Acid Synthase specific rabbit monoclonal antibody (1:1000 dilution, C20G5, Cell Signaling), SOD1 specific mouse monoclonal antibody (1:1000 dilution, 71G8, Cell Signaling), UCP3 specific rabbit monoclonal antibody (1:1000 dilution, D6J8K, Cell Signaling), TNF alpha specific rabbit polyclonal antibody (1:2000 dilution, AB6671, Abcam), Interferon gamma specific rabbit monoclonal antibody (1:1000 dilution, EPR1108, Abcam), were used as primary antisera. Horseradish peroxidase-conjugated goat anti-mouse immunoglobulin (1:2000 dilution, Thermo Scientific) or horseradish peroxidase-conjugated goat anti-rabbit immunoglobulin (1:2000 dilution, Thermo Scientific) were used to detect specific protein bands (explained in Figure Legends) using a chemiluminescence system (Nagajyothi et al. 2014). GDI (1: 10000 dilution, 71–0300, and rabbit polyclonal, Invitrogen, CA) and a secondary antibody horseradish peroxidase conjugated goat anti-rabbit (1:2000 dilution, Amersham Biosciences) was used to normalize protein loading.

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Manufacturer protocol

Download the product protocol from Abcam for Anti-ABCA1 antibody [AB.H10] (ab18180) below.

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