Proteins were isolated from snap frozen brown adipose tissue (BAT) by lysing tissues with RIPA buffer supplemented with protease inhibitors, after which the protein lysate was collected. Western blot was performed loading 4 µL of 0.2 µg/mL lysate on the Wes apparatus (ProteinSimple, Wiesbaden, Germany), according to the manufacturer’s protocol. The following primary and secondary antibodies were used: UCP1 (Sigma-Aldrich, U6382, 1:20), GAPDH (Santa Cruz Biotechnology, sc25778, 1:50), HRP anti-rabbit (ProteinSimple, DM001) and HRP anti-sheep (Abcam, Amsterdam, the Netherlands, ab6900, 1:100).
Dm001
Manufactured by Abcam
DM001 is a laboratory equipment product. It serves a core function in a laboratory setting, but a detailed description is not available at this time while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using dm001
1
Western Blot Analysis of UCP1 in BAT
2
Quantitative Western Blot Analysis
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Western blot was performed using the Wes apparatus (ProteinSimple), according to the manufacturer's protocol. For WAT and liver samples, 2.0 and 0.8 mg/mL lysate was used, respectively. The Wes apparatus allows simultaneous detection of multiple proteins and therefore loading controls may be identical between different figures. The following primary and secondary antibodies were used: 11BHSD1 (in house antibody University of Edinburgh, 1:10), FKBP5 (AB_2103136, R&D systems, AF4094, 1:70), GAPDH (AB_10167668, Santa Cruz Biotechnology, sc25778, 1:50), GR (AB_2631286, Cell Signaling, 12401, 1:10), HPR anti-goat (ProteinSimple, DM006), HRP antirabbit (ProteinSimple, DM001) and HRP anti-sheep (AB_955452, Abcam, ab6900, 1:100).
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