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|
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 blot analysis of extracts from various cell lines using HSL Antibody #4107.
Western blot analysis of extracts from differentiated NIH/3T3-L1 cells treated with isoproterenol, using Phospho-HSL (Ser660) Antibody #4126.
Western blot analysis of extracts from differentiated 3T3-L1 cells treated with forskolin, oligomycin or λ-phosphatase, using Phospho-HSL (Ser565) Antibody #4137.
Western blot analysis of extracts from differentiated 3T3-L1 cells treated with isoproterenol or λ-phosphatase, using Phospho-HSL (Ser563) Antibody #4139.
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.
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).
- Greenberg, A.S. et al. (1991) J Biol Chem 266, 11341-6.
- Brasaemle, D.L. (2007) J Lipid Res 48, 2547-59.
- Ducharme, N.A. and Bickel, P.E. (2008) Endocrinology 149, 942-9.
- Egan, J.J. et al. (1990) J Biol Chem 265, 18769-75.
- Brasaemle, D.L. et al. (2009) Mol Cell Biochem 326, 15-21.
- Degerman, E. et al. (1990) Proc Natl Acad Sci U S A 87, 533-7.
- Anthonsen, M.W. et al. (1998) J Biol Chem 273, 215-21.
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For Research Use Only. Not For Use In Diagnostic Procedures.