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8334
Lipolysis Activation Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Lipolysis Activation Antibody Sampler Kit #8334

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Lipolysis Activation Antibody Sampler Kit: Image 1

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).

Lipolysis Activation Antibody Sampler Kit: Image 2

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).

Lipolysis Activation Antibody Sampler Kit: Image 3

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).

Lipolysis Activation Antibody Sampler Kit: Image 4

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

Lipolysis Activation Antibody Sampler Kit: Image 5

Confocal immunofluorescent analysis of frozen mouse brown adipose tissue using Perilipin-1 (D1D8) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Lipolysis Activation Antibody Sampler Kit: 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.

Lipolysis Activation Antibody Sampler Kit: Image 7

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

Lipolysis Activation Antibody Sampler Kit: Image 8

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

Lipolysis Activation Antibody Sampler Kit: Image 9

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

Lipolysis Activation Antibody Sampler Kit: Image 10

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

Lipolysis Activation Antibody Sampler Kit: Image 11

Immunohistochemical analysis of paraffin-embedded breast carcinoma using Perilipin-1 (D1D8) XP® Rabbit mAb. Note specific staining of adipocytes.

Lipolysis Activation Antibody Sampler Kit: Image 12

Immunohistochemical analysis of parafin-embedded mouse brown fat using Perilipin-1 (D1D8) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).

Lipolysis Activation Antibody Sampler Kit: Image 13

Immunohistochemical analysis of parafin-embedded mouse kidney with staining of adjacent adipose tissue using Perilipin-1 (D1D8) XP® Rabbit mAb.

Lipolysis Activation Antibody Sampler Kit: Image 14

Immunohistochemical analysis of parafin-embedded mouse prostate with staining of adjacent adipose tissue using Perilipin-1 (D1D8) XP® Rabbit mAb.

Lipolysis Activation Antibody Sampler Kit: Image 15

Immunohistochemical analysis of parafin-embedded mouse testis with staining of adjacent adipose tissue using Perilipin-1 (D1D8) XP® Rabbit mAb.

Lipolysis Activation Antibody Sampler Kit: Image 16

Western blot analysis of extracts from human pre-adipocytes and adipocytes using Perilipin-1 (D1D8) XP® Rabbit mAb (upper) and β-Actin Antibody #4967 (lower).

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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-1 (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 

Product 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 two western mini-blot experiments with each primary antibody.

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.

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. USA 87, 533-37.
  7. Anthonsen, M.W. et al. (1998) J. Biol. Chem. 273, 215-21.

Pathways & Proteins

Explore pathways + proteins related to this product.

Limited Uses

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Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST's products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST's Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

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