3T3L1 adipocytes or 100–150 mgs of ground eWAT were lysed in ice-cold RIPA buffer with 1× protease inhibitor (Sigma-Aldrich). 30 μg of total protein was separated on a 12% SDS-PAGE gel for BCKDH blots or 20 ugs of total protein was separated on a 4–20% SDS-PAGE gel (mini-protean TGX gels, Bio-Rad) for CrAT and FASN blots. The proteins were transferred to a nitrocellulose membrane and immunoblotted with anti-Bckdha (Novus Biologicals NBP1–79616) (1:1,000 dilution), anti-Beta-Actin (Cell Signaling 4970S) (1:5,000), anti-GAPDH (Cell signaling 5174S), anti-CrAT (Novus NBP2–15999), anti-FASN (Proteintech 10624–2-AP). Specific signal was detected with horseradish peroxidase-conjugated secondary antibody goat anti-rabbit (1:2,500 – 1:10,000) using SuperSignal West Pico Chemiluminescent Substrate (Thermo Scientific) and developed using Blue Devil Autoradiography film (Genesee Scientific) or BioRad Chemidoc XRS+.
Anti crat
Manufactured by Novus Biologicals
Anti-CrAT is a primary antibody that targets the Carnitine O-acetyltransferase (CrAT) protein. CrAT is an enzyme involved in the transport of acetyl-CoA into the mitochondria for energy production. This antibody can be used to detect and quantify the CrAT protein in various biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Protease inhibitor, by Merck Group (2 mentions)
Anti fasn, by Proteintech (2 mentions)
Anti bckdha, by Novus Biologicals (2 mentions)
Blue devil autoradiography film, by Genesee Scientific (2 mentions)
Anti β actin, by Cell Signaling Technology (2 mentions)
Supersignal west pico chemiluminescent substrate, by Thermo Fisher Scientific (2 mentions)
Anti gapdh, by Cell Signaling Technology (2 mentions)
Mini protean tgx gel, by Bio-Rad (2 mentions)
2 protocols using anti crat
1
Quantification of Adipocyte Proteins
2
Quantification of Adipocyte Proteins
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3T3L1 adipocytes or 100–150 mgs of ground eWAT were lysed in ice-cold RIPA buffer with 1× protease inhibitor (Sigma-Aldrich). 30 μg of total protein was separated on a 12% SDS-PAGE gel for BCKDH blots or 20 ugs of total protein was separated on a 4–20% SDS-PAGE gel (mini-protean TGX gels, Bio-Rad) for CrAT and FASN blots. The proteins were transferred to a nitrocellulose membrane and immunoblotted with anti-Bckdha (Novus Biologicals NBP1–79616) (1:1,000 dilution), anti-Beta-Actin (Cell Signaling 4970S) (1:5,000), anti-GAPDH (Cell signaling 5174S), anti-CrAT (Novus NBP2–15999), anti-FASN (Proteintech 10624–2-AP). Specific signal was detected with horseradish peroxidase-conjugated secondary antibody goat anti-rabbit (1:2,500 – 1:10,000) using SuperSignal West Pico Chemiluminescent Substrate (Thermo Scientific) and developed using Blue Devil Autoradiography film (Genesee Scientific) or BioRad Chemidoc XRS+.
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