Cell Signaling Technology

Product Pathways - Metabolism

Phospho-HSL (Ser563) Antibody #4139

Applications Reactivity Sensitivity MW (kDa) Source
W IF-IC M (H) (R) Endogenous 81, 83 Rabbit

Applications Key:  W=Western Blotting  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  M=Mouse  R=Rat
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

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

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser552 of human HSL (equivalent to Ser563 of rat HSL). Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from differentiated 3T3-L1 cells treated with isoproterenol or lambda protein phosphatase, using Phospho-HSL (Ser563) Antibody.

IF-IC

IF-IC

Confocal immunofluorescent analysis of 3T3-L1 adipocytes, isoproterenol-treated (left) or phosphatase-treated (right), labeled with Phospho-HSL (Ser563) Antibody (red). Lipid droplets have been labeled with BODIPY 493/503 (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Background

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.

Application References

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Companion Products

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

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