Protein extracts of cells were prepared using RIPA buffer containing protease inhibitor cocktail, and protein concentrations were measured using a BCA kit (Pierce). One hundred micrograms of protein was resolved by electrophoresis in 8% SDS‐PAGE gel and blotted to PVDF membranes (Invitrogen). Membranes were incubated with indicated primary antibodies overnight at 4°C and then with an anti‐rabbit HRP‐conjugated secondary antibody (1:3000; Santa Cruz Biotechnology) for 2 hours at room temperature. The primary antibodies used in this study were rabbit anti‐FAS (1:1000; Cell signaling), rabbit anti‐SCD1 (1:500; Cell signaling), rabbit anti‐FADS2 (1:1000; Abcam), rabbit anti‐COX2 (1:1000; Abcam), and anti‐β‐actin (1:2000; Santa Cruz Biotechnology) as a loading control. HRP activity was measured using Supersignal West Pico Chemiluminescent Substrate kit (Thermo Fisher Scientific).
Rabbit anti fas
Manufactured by Cell Signaling Technology
Sourced in United States
Rabbit anti‐FAS is a primary antibody that recognizes the FAS (CD95/APO-1) protein. FAS is a cell surface receptor that plays a key role in the initiation of apoptosis (programmed cell death). This antibody can be used to detect the expression and activation of FAS in various cell and tissue samples.
Automatically generated - may contain errors
Lab products found in correlation
Supersignal west pico chemiluminescent substrate kit, by Thermo Fisher Scientific (1 mentions)
Hrp conjugated anti rabbit secondary antibody, by Santa Cruz Biotechnology (1 mentions)
Pvdf membrane, by Thermo Fisher Scientific (1 mentions)
Rabbit anti cox 2, by Abcam (1 mentions)
Anti β actin, by Santa Cruz Biotechnology (1 mentions)
Rabbit anti fads2, by Abcam (1 mentions)
Rabbit anti scd1, by Cell Signaling Technology (1 mentions)
Goat serum, by Merck Group (1 mentions)
Extravidine, by Merck Group (1 mentions)
Anti srebp1, by Abcam (1 mentions)
Anti pparg, by Thermo Fisher Scientific (1 mentions)
2 protocols using rabbit anti fas
1
Western Blot Analysis of Lipid Metabolism Proteins
2
Immunohistochemistry on Paraffin Sections
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Immunohistochemistry on paraffin sections (3 µm) was performed as previously described (Zellmer et al., 2009 (link)). The sections were boiled (3 × 5 min) in buffer (0.01 M sodium citrate, pH 6.0) and the slides were incubated for 30 min in 5% goat serum (Sigma-Aldrich, Germany) to block nonspecific binding. The following antibodies were used: rabbit anti-FAS (1:400, Cell Signaling Technology, USA), rabbit anti-Cre (1:4000; Abcam, Cambridge), anti-GLI3 (1:1000; GeneTex, USA), anti-GLI1 (1:1000; GeneTex, USA), anti-SREBP1 (1:200; Abcam, Cambridge) anti-PPARG (1:250, ThermoFisher Scientific, Germany), biotinylated goat anti-rabbit IgG (Millipore, Germany) and Extravidine (Sigma-Aldrich, Germany). POD and counterstaining with hematoxylin were performed as previously described (Zellmer et al., 2009 (link)).
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