Brown adipose tissues were lysed in RIPA buffer with Complete Protease Inhibitor Cocktail (Roche Inc., Indianapolis, IN). Protein concentration was determined with BCA protein assay kit (Pierce, Rockford, IL). Twenty microgram of protein of each sample was separated by SDS-PAGE, and electro-transferred to nitrocellulose membrane for immunoblot analyses. The following antibodies were used: anti-p-PKA (Tyr197) (Cell Signaling, Danvers, MA, 4781S, 1:1000), anti-t-PKA (Cell Signaling, Danvers, MA, 4782, 1:1000), anti-p-HSL (Ser563) (Cell Signaling, Danvers, MA, 4139S, 1:1000), anti-t-HSL (Cell Signaling, Danvers, MA, 4107, 1:1000), anti-p-Creb (Ser133) (Cell Signaling, Danvers, MA, 9191S, 1:1000), anti-t-Creb (Cell Signaling, Danvers, MA, 9197, 1:1000), p-AMPKα (T172) (Cell Signaling, Danvers, MA, 2535L, 1:1000), anti t-AMPK (Cell Signaling, Danvers, MA, 2603S, 1:1000), anti-UCP1 (ABCAM, Cambridge, MA Ab10983, 1:10000), anti-β-actin (Cell Signaling, Danvers, MA, 4967S, 1:1000), HRP-conjugated anti-mouse (GE Healthcare UK Limited, 1:10,000), and anti-rabbit (GE Healthcare UK Limited, 1:10,000). The SuperSignal West Pico Chemiluminescent kit (Pierce, Rockford, IL) was used as substrates.
Anti t hsl
Manufactured by Cell Signaling Technology
Anti-t-HSL is a primary antibody that recognizes the phosphorylated form of the hormone-sensitive lipase (HSL) enzyme. HSL is a key regulator of lipolysis, the process of breaking down stored fats. The Anti-t-HSL antibody can be used to detect the activated form of HSL, which is phosphorylated at specific threonine residues.
Automatically generated - may contain errors
Lab products found in correlation
Anti β actin, by Cell Signaling Technology (2 mentions)
Anti p hsl ser563, by Cell Signaling Technology (1 mentions)
Anti tcreb, by Cell Signaling Technology (1 mentions)
P ampkαt172, by Cell Signaling Technology (1 mentions)
Anti t ampk, by Cell Signaling Technology (1 mentions)
Anti ucp1, by Abcam (1 mentions)
Complete protease inhibitor cocktail, by Roche (1 mentions)
Anti p creb ser133, by Cell Signaling Technology (1 mentions)
Anti atgl, by Cell Signaling Technology (1 mentions)
Anti α tubulin, by Abcam (1 mentions)
Pparγ, by Cell Signaling Technology (1 mentions)
Anti p hsl, by Cell Signaling Technology (1 mentions)
Anti pgc 1α, by Abcam (1 mentions)
2 protocols using anti t hsl
1
Immunoblotting of Brown Adipose Tissue
2
Western Blot Analysis of Adipogenic Proteins
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Western blot analysis was conducted as previously reported.15 (link) The target protein was detected using primary antibodies as follows: anti-proliferator-activated receptor gamma (PPAR-γ) (1:1000, Cell Signaling Technology), anti-PGC-1α (1:1000, Abcam), anti-ATGL (1:1000, Cell Signaling Technology), anti-pHSL (1:1000, Cell Signaling Technology), anti-tHSL (1:1000, Cell Signaling Technology), anti-α-tubulin (1:1000, Abcam) and anti-β-actin (1:2000, Cell Signaling Technology).
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