Product Pathways - Glucose Metabolism
Phospho-HSL (Ser660) Antibody #4126
PhosphoSitePlus® protein, site, and accession data: HSL
| Applications | Reactivity | Sensitivity | MW (kDa) | Source |
|---|---|---|---|---|
| W IF-IC | M R | Endogenous | 81, 83 | Rabbit |
Applications Key:
W=Western Blotting
IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:
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 (Ser660) Antibody detects endogenous levels of HSL only when phosphorylated at Ser660 by PKA. This antibody does not cross-react with Ser563, Ser565 or Ser659 phosphorylated HSL.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser651 of mouse HSL (equivalent to Ser660 of rat HSL). Antibodies are purified by protein A and peptide affinity chromatography.
Western Blotting
Western blot analysis of extracts from differentiated NIH/3T3-L1 cells treated with isoproterenol, using Phospho-HSL (Ser660) Antibody.
IF-IC
Confocal immunofluorescent analysis of 3T3-L1 adipocytes, isoproterenol-treated (left) or untreated (right), labeled with Phospho-HSL (Ser660) 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).
- Degerman, E. et al. (1990) Proc. Natl. Acad. Sci. USA 87, 533-537.
- Anthonsen, M.W. et al. (1998) J. Biol. Chem. 273, 215-221.
- Garton, A.J. and Yeaman, S.J. (1990) Eur. J. Biochem. 191, 245-250.
- Greenberg, A.S. et al. (2001) J. Biol. Chem. 276, 45456-45461.
Application References
- Aboulaich, N. et al. (2011) Diabetes 60, 757-65. Applications: Western Blotting
- Choi, S.M. et al. (2010) Mol Cell Biol , . Applications: Western Blotting
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
Companion Products
- 4107 HSL Antibody
- 4137 Phospho-HSL (Ser565) Antibody
- 4139 Phospho-HSL (Ser563) Antibody
- 9621 Phospho-(Ser/Thr) PKA Substrate Antibody
- 9624 Phospho-PKA Substrate (RRXS*/T*) (100G7E) Rabbit mAb
- 2535 Phospho-AMPKα (Thr172) (40H9) Rabbit mAb
- 2603 AMPKα (23A3) Rabbit mAb
- 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
