Revision 4

#8334Store at -20C

1 Kit

(5 x 20 microliters)

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
Phospho-HSL (Ser563) Antibody 4139 20 µl 81, 83 kDa Rabbit 
Phospho-HSL (Ser565) Antibody 4137 20 µl 81, 83 kDa Rabbit 
Perilipin-1 (D1D8) XP® Rabbit mAb 9349 20 µl 62 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 
HSL (D6W5S) XP® Rabbit mAb 18381 20 µl 81, 83 kDa Rabbit IgG
Phospho-HSL (Ser660) Antibody 45804 20 µl 81, 83 kDa Rabbit 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

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.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

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.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    使用に関する制限

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    研究専用 (For Research Use Only) またはこれに類似する表示がされた製品は、 いかなる目的についても FDA または外国もしくは国内のその他の規制機関により承認、認可または許可を受けていません。 お客様は製品を診断もしくは治療目的で使用してはならず、また、製品に表示された内容に違反する方法で使用してはなりません。 CST が販売または使用許諾する製品は、エンドユーザーであるお客様に対し、使途を研究および開発のみに限定して提供されるものです。 診断、予防もしくは治療目的で製品を使用することまたは製品を再販売 (単独であるか他の製品等の一部であるかを問いません) もしくはその他の商業的利用の目的で購入することについては、CST から別途許諾を得る必要があります。 お客様は以下の事項を遵守しなければなりません。(a) CST の製品 (単独であるか他の資材と一緒であるかを問いません) を販売、使用許諾、貸与、寄付もしくはその他の態様で第三者に譲渡したり使用させたりしてはなりません。また、商用の製品を製造するために CST の製品を使用してはなりません。(b) 複製、改変、リバースエンジニアリング、逆コンパイル、 分解または他の方法により製品の構造または技術を解明しようとしてはなりません。また、 CST の製品またはサービスと競合する製品またはサービスを開発する目的で CST の製品を使用してはなりません。(c) CST の製品の商標、商号、ロゴ、特許または著作権に関する通知または表示を除去したり改変したりしてはなりません。(d) CST の製品をCST 製品販売条件(CST’s Product Terms of Sale) および該当する書面のみに従って使用しなければなりません。(e) CST の製品に関連してお客様が使用する第三者の製品またはサービスに関する使用許諾条件、 サービス提供条件またはこれに類する合意事項を遵守しなければなりません。

    Revision 4
    #8334

    Lipolysis Activation Antibody Sampler Kit

    Lipolysis Activation Antibody Sampler Kit: Image 1 Expand Image
    Confocal immunofluorescent analysis of 10-day differentiated 3T3-L1 cells (left, positive) or undifferentiated 3T3-L1 cells (right, negative) using HSL (D6W5S) XP® Rabbit mAb (green), Perilipin-1 (D1D8) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) #52975 (red), and DAPI #4083 (blue).
    Lipolysis Activation Antibody Sampler Kit: Image 2 Expand Image
    Immunoprecipitation of perilipin-1 protein from 3T3-L1 cell extracts differentiated for 4 days. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Perilipin-1 (D1D8) XP® Rabbit mAb. Mouse Anti-rabbit IgG (Light-Chain Specific) (L57A3) mAb #3677 and Anti-mouse IgG, HRP-linked Antibody #7076 were used as the secondary antibodies.
    Lipolysis Activation Antibody Sampler Kit: Image 3 Expand Image
    Western blot analysis of extracts from undifferentiated and differentiated 3T3-L1 cells using HSL (D6W5S) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
    Lipolysis Activation Antibody Sampler Kit: Image 4 Expand Image
    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 5 Expand Image
    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 6 Expand Image
    Western blot analysis of extracts from 3T3-L1 cells, undifferentiated (lane 1) or differentiated 4 days into adipocytes and treated with isoproterenol (10 μM, 20 min; lane 2), using Phospho-HSL (Ser660) Antibody (upper) or α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower).
    Lipolysis Activation Antibody Sampler Kit: Image 7 Expand Image
    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 8 Expand Image
    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).
    Lipolysis Activation Antibody Sampler Kit: Image 9 Expand Image
    Western blot analysis of extracts from human pre-adipocytes and differentiated human adipocytes using HSL (D6W5S) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
    Lipolysis Activation Antibody Sampler Kit: Image 10 Expand Image
    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 11 Expand Image
    Western blot analysis of extracts from 3T3-L1 cells, undifferentiated (lane 1) or differentiated 4 days into adipocytes and treated with isoproterenol (10 μM, 20 min; lane 2), using Phospho-HSL (Ser660) Antibody. Phospho-specificity was verified by incubating the antibody without peptide (left), with HSL (Ser660) non-phosphopeptide (middle), or with HSL (Ser660) phosphopeptide (right) prior to incubation with the membrane.
    Lipolysis Activation Antibody Sampler Kit: Image 12 Expand Image
    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 13 Expand Image
    Immunoprecipitation of HSL from differentiated human adipocyte extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is HSL (D6W5S) XP® Rabbit mAb. Western blot analysis was performed usingHSL (D6W5S) XP® Rabbit mAb.
    Lipolysis Activation Antibody Sampler Kit: Image 14 Expand Image
    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 15 Expand Image
    Confocal immunofluorescent analysis of differentiated human adipocytes (left) or human pre-adipocytes (right) using HSL (D6W5S) XP® Rabbit mAb (green). Actin filaments were labeled with DyLight® 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Lipolysis Activation Antibody Sampler Kit: Image 16 Expand Image
    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 17 Expand Image
    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 18 Expand Image
    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 19 Expand Image
    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 20 Expand Image
    Confocal immunofluorescent analysis of 8-day differentiated 3T3-L1 cells, using Perilipin (D1D8) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).