Cell Signaling Technology

Product Pathways - Metabolism

Lipolysis Activation Antibody Sampler Kit #8334

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
HSL Antibody #4107 40 µl W IP IF-IC M (H) 81, 83 Rabbit
Phospho-HSL (Ser563) Antibody #4139 40 µl W IF-IC M (H) (R) 81, 83 Rabbit
Phospho-HSL (Ser565) Antibody #4137 40 µl W IF-IC M (H) (R) 81, 83 Rabbit
Phospho-HSL (Ser660) Antibody #4126 40 µl W IF-IC M R 81, 83 Rabbit
Perilipin (D1D8) XP® Rabbit mAb #9349 40 µl W IP IHC-P IF-F IF-IC H M 62 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl Goat

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-F=Immunofluorescence (Frozen)  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  M=Mouse  R=Rat
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

Each antibody in the Lipolysis Activation Antibody Sampler Kit detects endogenous levels of the respective target protein. Phospho-HSL (Ser563) Antibody does not cross-react with Ser565 phosphorylated HSL. Phospho-HSL (Ser565) Antibody does not cross-react with Ser563 phosphorylated HSL. Phospho-HSL (Ser660) Antibody does not cross-react with Ser563, Ser565, or Ser659 phosphorylated HSL.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using HSL Antibody #4107.

Western Blotting

Western Blotting

Western blot analysis of extracts from differentiated NIH/3T3-L1 cells treated with isoproterenol, using Phospho-HSL (Ser660) Antibody #4126.

Western Blotting

Western Blotting

Western blot analysis of extracts from differentiated 3T3-L1 cells treated with forskolin, oligomycin or λ-phosphatase, using Phospho-HSL (Ser565) Antibody #4137.


Western Blotting

Western Blotting

Western blot analysis of extracts from differentiated 3T3-L1 cells treated with isoproterenol or λ-phosphatase, using Phospho-HSL (Ser563) Antibody #4139.

Western Blotting

Western Blotting

Western blot analysis of extracts from human pre-adipocytes and adipocytes using Perilipin (D1D8) XP® Rabbit mAb #9349 (upper) and β-Actin Antibody #4967 (lower).

Description

The Lipolysis Activation Antibody Sampler Kit provides an economical means to evaluate the activation status of multiple members of the lipolysis pathway, including phosphorylated HSL and perilipin. The kit includes enough antibody to perform four western mini-blot experiments with each primary antibody.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser552 of human HSL (equivalent to Ser563 of rat HSL), Ser554 of human HSL (equivalent to Ser565 of rat HSL), Ser651 of mouse HSL (equivalent to Ser660 of rat HSL), or the sequence of human HSL. Polyclonal antibodies are purified by protein A and peptide affinity chromatography. Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues around Ile419 of human perilipin protein.

Background

Triacylglycerol is stored in lipid droplets as a primary energy reserve. During lipolysis, triacylglycerols in adipocytes are hydrolyzed into free fatty acids and glycerol. Perilipin, localized at the periphery of lipid droplets, serves as a protective coating against lipases (1-3). Evidence suggests that PKA regulates lipolysis by phosphorylating perilipin and hormone-sensitive lipase (HSL) (1,2,4,5). Phosphorylation of perilipin results in the conformational change that exposes lipid droplets to endogenous lipases, such as HSL (2). Phosphorylation of HSL at Ser563, Ser659, and Ser660 by PKA stimulates HSL activity, which in turn catalyzes the hydrolysis of triacylglycerol (6,7).

  1. Greenberg, A.S. et al. (1991) J Biol Chem 266, 11341-6.
  2. Brasaemle, D.L. (2007) J Lipid Res 48, 2547-59.
  3. Ducharme, N.A. and Bickel, P.E. (2008) Endocrinology 149, 942-9.
  4. Egan, J.J. et al. (1990) J Biol Chem 265, 18769-75.
  5. Brasaemle, D.L. et al. (2009) Mol Cell Biochem 326, 15-21.
  6. Degerman, E. et al. (1990) Proc Natl Acad Sci U S A 87, 533-7.
  7. Anthonsen, M.W. et al. (1998) J Biol Chem 273, 215-21.

Application References

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

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