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8334
Lipolysis Activation Antibody Sampler Kit

Lipolysis Activation Antibody Sampler Kit #8334

Western Blotting Image 1

Western blot analysis of extracts from NIH/3T3 and 3T3-L1 cells using HSL Antibody.

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Western Blotting Image 2

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

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Western Blotting Image 3

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

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Western Blotting Image 4

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

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Western Blotting Image 5

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

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Western Blotting Image 6

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

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IF-IC Image 7

Confocal immunofluorescent analysis of 3T3-L1 adipocytes using HSL Antibody (red). Lipid droplets have been labeled with BODIPY 493/503 (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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IF-IC Image 8

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).

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IF-IC Image 9

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).

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IHC-P (paraffin) Image 10

Immunohistochemical analysis of paraffin-embedded breast carcinoma using Perilipin (D1D8) XP® Rabbit mAb. Note specific staining of adipocytes.

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IHC-P (paraffin) Image 11

Immunohistochemical analysis of parafin-embedded mouse brown fat using Perilipin (D1D8) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).

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IF-IC Image 12

Confocal immunofluorescent analysis of 8-day differentiated 3T3-L1 cells, using Perilipin (D1D8) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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IF-IC Image 13

Confocal immunofluorescent analysis of frozen mouse brown adipose tissue using Perilipin (D1D8) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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Product Includes Quantity Applications Reactivity MW(kDa) Isotype
HSL Antibody 4107 20 µl
  • WB
  • IP
  • IF
H M 81, 83 Rabbit 
Phospho-HSL (Ser563) Antibody 4139 20 µl
  • WB
  • IF
M 81, 83 Rabbit 
Phospho-HSL (Ser565) Antibody 4137 20 µl
  • WB
  • IF
M 81, 83 Rabbit 
Phospho-HSL (Ser660) Antibody 4126 20 µl
  • WB
M R 81, 83 Rabbit 
Perilipin (D1D8) XP® Rabbit mAb 9349 20 µl
  • WB
  • IP
  • IHC
  • IF
H M 62 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

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 two western mini-blot experiments with each primary antibody.

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.

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).

  1. Degerman, E. et al. (1990) Proc. Natl. Acad. Sci. USA 87, 533-37.
  2. Anthonsen, M.W. et al. (1998) J. Biol. Chem. 273, 215-21.
  3. Greenberg, A.S. et al. (1991) J Biol Chem 266, 11341-6.
  4. Brasaemle, D.L. (2007) J Lipid Res 48, 2547-59.
  5. Ducharme, N.A. and Bickel, P.E. (2008) Endocrinology 149, 942-9.
  6. Egan, J.J. et al. (1990) J Biol Chem 265, 18769-75.
  7. Brasaemle, D.L. et al. (2009) Mol Cell Biochem 326, 15-21.
Entrez-Gene Id
3991 , 5346
Swiss-Prot Acc.
Q05469 , O60240
For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

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