Phospho-HSL (Ser565) Antibody

4137:

Immunofluorescence , Western Blotting

Phospho-HSL (Ser565) Antibody detects endogenous levels of HSL only when phosphorylated at Ser565 by AMPK. This antibody does not cross-react with Ser563 phosphorylated HSL.

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser554 of human HSL (Ser565 of rat HSL). Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western blot analysis of extracts from differentiated 3T3-L1 cells treated with forskolin, oligomycin or lambda protein phosphatase, using Phospho-HSL (Ser565) Antibody.

IF-IC

Confocal immunofluorescent analysis of 3T3-L1 cells, untreated (left) or phosphatase-treated (right), labeled with Phospho-HSL (Ser565) Antibody (red) showing cytoplasmic localization in differentiated cells. Lipid droplets have been labeled with BODIPY® 493/503 (green). Blue pseudocolor = DRAQ5^® #4084 (fluorescent DNA dye).

HSL (hormone-sensitive lipase) catalyzes the hydrolysis of triacylglycerol, the rate-limiting step in lipolysis. Lipolytic stimuli activate adenylyl cyclase and thus increase intracellular cAMP levels, which in turn activate protein kinase A (PKA). PKA phosphorylates HSL at Ser563, Ser659 and Ser660, which stimulates HSL activity (1,2). In contrast, AMPK phosphorylates HSL at Ser565, which reduces HSL phosphorylation at Ser563 by PKA and inhibits HSL activity (2,3). Recent work indicates that phosphorylation at Ser600 by p44/42 MAPKs also enhances the enzymatic activity of HSL (4).

1. Degerman, E. et al. (1990) Proc. Natl. Acad. Sci. USA 87, 533-537.
2. Anthonsen, M.W. et al. (1998) J. Biol. Chem. 273, 215-221.
3. Garton, A.J. and Yeaman, S.J. (1990) Eur. J. Biochem. 191, 245-250.
4. Greenberg, A.S. et al. (2001) J. Biol. Chem. 276, 45456-45461.

• Loo, L.H. et al. (2009) J Cell Biol 187, 375-84. Applications: IF-IC (In Cells)

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For Research Use Only. Not For Use In Diagnostic Procedures.