Revision 1

#8335Store at -20C

1 个试剂盒

(8 x 20 microliters)

Cell Signaling Technology

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Support: 877-678-TECH (8324)

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For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
AceCS1 (D19C6) Rabbit mAb 3658 20 µl 78 kDa Rabbit IgG
Phospho-Acetyl-CoA Carboxylase (Ser79) (D7D11) Rabbit mAb 11818 20 µl 280 kDa Rabbit IgG
Acetyl-CoA Carboxylase (C83B10) Rabbit mAb 3676 20 µl 280 kDa Rabbit IgG
ATP-Citrate Lyase Antibody 4332 20 µl 125 kDa Rabbit 
Phospho-ATP-Citrate Lyase (Ser455) Antibody 4331 20 µl 125 kDa Rabbit 
Fatty Acid Synthase (C20G5) Rabbit mAb 3180 20 µl 273 kDa Rabbit IgG
Lipin 1 (D2W9G) Rabbit mAb 14906 20 µl 130 kDa Rabbit IgG
ACSL1 (D2H5) Rabbit mAb 9189 20 µl 78 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

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

Description

The Fatty Acid and Lipid Metabolism Antibody Sampler Kit provides an economical means to evaluate key proteins involved in fatty acid and lipid metabolism. This kit includes enough primary antibody to perform two western miniblot 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

The processes of fatty acid and lipid metabolism are vital for cellular nutrient and energy maintenance. Cytoplasmic acetyl-CoA synthetase (AceCS1) catalyzes the conversion of acetate and CoA to acetyl-CoA. Acetyl-CoA synthesized by AceCS1 is used for fatty acid and lipid biosynthesis (1,2). Acetyl-CoA carboxylase (ACC) catalyzes the pivotal step of the fatty acid synthesis pathway. Phosphorylation by AMPK at Ser79 or by PKA at Ser1200 inhibits the enzymatic activity of ACC (3). Mammalian long-chain acyl-CoA synthetase (ACSL) catalyzes the ligation of the fatty acid to CoA to form fatty acyl-CoA in a two-step reaction (4). ATP-citrate lyase (ACL) is a homotetramer that catalyzes the formation of acetyl-CoA and oxaloacetate (OAA) in the cytosol, which is the key step for the biosynthesis of fatty acids, cholesterol, and acetylcholine, as well as for glucogenesis (5). Phosphorylation of ACL at Ser455 abolishes the homotropic allosteric regulation by citrate and enhances the catalytic activity of the enzyme (6). Fatty acid synthase (FASN) catalyzes the synthesis of long-chain fatty acids from acetyl-CoA and malonyl-CoA (7). Lipin 1 plays a role in lipid metabolism in various tissues and cell types including liver, muscle, adipose tissues, and neuronal cell lines (8-10). It has dual functions at the molecular level: Lipin 1 serves as a transcriptional coactivator in the liver and a phosphatidate phosphatase in triglyceride and phospholipid biosynthesis pathways (11).

  1. Ikeda, Y. et al. (2001) J Biol Chem 276, 34259-69.
  2. Luong, A. et al. (2000) J Biol Chem 275, 26458-66.
  3. Ha, J. et al. (1994) J Biol Chem 269, 22162-8.
  4. Mashek, D.G. et al. (2004) J Lipid Res 45, 1958-61.
  5. Towle, H.C. et al. (1997) Annu Rev Nutr 17, 405-33.
  6. Potapova, I.A. et al. (2000) Biochemistry 39, 1169-79.
  7. Katsurada, A. et al. (1990) Eur J Biochem 190, 427-33.
  8. Finck, B.N. et al. (2006) Cell Metab 4, 199-210.
  9. Phan, J. and Reue, K. (2005) Cell Metab 1, 73-83.
  10. Verheijen, M.H. et al. (2003) Genes Dev 17, 2450-64.
  11. Reue, K. and Zhang, P. (2008) FEBS Lett 582, 90-6.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

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    Orders: 877-616-CELL (2355)•[email protected]• Support: 877-678-TECH (8324)•[email protected]• Web: cellsignal.com
    仅供研究使用。不得用于诊断流程。
    Revision 1
    #8335

    Fatty Acid and Lipid Metabolism Antibody Sampler Kit

    Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 1 Expand Image
    Simple Western™️ analysis of lysates (1mg/mL) from 3T3 cells using Acetyl-CoA Carboxylase (C83B10) Rabbit mAb #3676. 虚拟泳道式图像（左图）显示一抗稀释比例为 1:10 和 1:50 时的靶标条带（如图所示）。对应的电泳图（右图）为一抗稀释比例在 1:10（蓝线）和 1:50（绿线）时沿毛细血管内分子量的化学发光结果。在还原条件下，使用 66-440 kDa 分离模块在 ProteinSimple（BioTechne 品牌）的 Jess™ Simple Western 仪器上进行该实验。
    Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 2 Expand Image
    使用 Phospho-Acetyl-CoA Carboxylase (Ser79) (D7D11) Rabbit mAb（上）或 Acetyl-CoA Carboxylase (C83B10) Rabbit mAb #3676（下）对未经或经寡霉素 #9996 处理（0.5 μM，30 分钟）的 SH-SY5Y 细胞提取物进行蛋白质印迹分析。磷酸化抗体的特异性可通过 λ 磷酸酶处理进行验证。
    Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 3 Expand Image
    使用 Lipin 1 (D2W9G) Rabbit mAb（上）和 GAPDH (D16H11) XP® Rabbit mAb #5174（下）对不同细胞系提取物进行蛋白质印迹分析。
    Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 4 Expand Image
    使用 Fatty Acid Synthase (C20G5) Rabbit mAb 对不同细胞类型的提取物进行蛋白质印迹分析。
    Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 5 Expand Image
    使用 AceCS1 (D19C6) Rabbit mAb 对不同细胞类型的提取物进行蛋白质印迹分析。
    Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 6 Expand Image
    使用 Acetyl-CoA Carboxylase (C83B10) Rabbit mAb 对不同细胞系细胞提取物进行蛋白质印迹分析。
    Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 7 Expand Image
    使用 Phospho-ATP-Citrate Lyase (Ser455) Antibody（上图）或 ATP-Citrate Lyase Antibody #4332（下图）对未经或经 PDGF 处理的指定时间的 NIH/3T3 细胞提取物进行蛋白质印迹分析。
    Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 8 Expand Image
    使用 ATP-Citrate Lyase Antibody 对 HeLa、NIH/3T3、C6 和 COS 细胞提取物进行蛋白质印迹分析。
    Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 9 Expand Image
    一抗与靶标蛋白结合之后，与偶联 HRP 的二抗形成复合体。添加 LumiGLO®，在酶催化分解期间发光。
    Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 10 Expand Image
    使用 ACSL1 (D2H5) Rabbit mAb 对不同细胞系提取物进行蛋白质印迹分析。
    Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 11 Expand Image
    Simple Western™ analysis of lysates (0.1 mg/mL) from Hela untreated cells using Phospho-Acetyl-CoA Carboxylase (Ser79) (D7D11) Rabbit mAb #11818. 虚拟泳道式图像（左图）显示一抗稀释比例为 1:10 和 1:50 时的单一靶标条带（如图所示）。对应的电泳图（右图）为一抗稀释比例在 1:10（蓝线）和 1:50（绿线）时沿毛细血管内分子量的化学发光结果。在还原条件下，使用 66-440 kDa 分离模块在 ProteinSimple（BioTechne 品牌）的 Jess™ Simple Western 仪器上进行该实验。
    Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 12 Expand Image
    使用 Lipin 1 (D2W9G) Rabbit mAb（红色）对 3T3-L1 脂肪细胞（分化，7天）进行共聚焦免疫荧光分析。用 BODIPY® 493/503（绿色）标记脂肪滴。蓝色伪彩 = DRAQ5® #4084（DNA 荧光染料）。
    Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 13 Expand Image
    使用 Fatty Acid Synthase (C20G5) Rabbit mAb 对石蜡包埋的人类乳腺癌细胞进行免疫组织化学分析。
    Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 14 Expand Image
    对 HeLa 细胞提取物的乙酰辅酶 A 羧化酶进行免疫沉淀。泳道 1 是 10% input，泳道 2 是 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900，并且泳道 3 是 Acetyl-CoA Carboxylase (C83B10) Rabbit mAb。使用 Acetyl-CoA Carboxylase (C83B10) Rabbit mAb 进行蛋白质印迹分析。
    Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 15 Expand Image
    使用 Phospho-Acetyl-CoA Carboxylase (Ser79) (D7D11) Rabbit mAb 对石蜡包埋的人乳腺癌细胞进行免疫组织化学分析。
    Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 16 Expand Image
    使用 Fatty Acid Synthase (C20G5) Rabbit mAb 对石蜡包埋的肝细胞癌细胞进行免疫组织化学分析。
    Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 17 Expand Image
    在对照肽（左）或 Acetyl-CoA Carboxylase (C83B10) Blocking Peptide #1062（右）存在的情况下，使用 Acetyl-CoA Carboxylase (C83B10) Rabbit mAb 对石蜡包埋的人乳腺癌细胞进行免疫组织化学分析。
    Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 18 Expand Image
    使用 Phospho-Acetyl-CoA Carboxylase (Ser79) (D7D11) Rabbit mAb 对石蜡包埋的小鼠肝脏细胞进行免疫组织化学分析。
    Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 19 Expand Image
    使用 Fatty Acid Synthase (C20G5) Rabbit mAb 对石蜡包埋的肺癌细胞进行免疫组织化学分析。
    Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 20 Expand Image
    使用 Acetyl-CoA Carboxylase (C83B10) Rabbit mAb 对石蜡包埋的人乳腺癌细胞进行免疫组织化学分析。
    Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 21 Expand Image
    使用 Phospho-Acetyl-CoA Carboxylase (Ser79) (D7D11) Rabbit mAb 对未经（左）或经 λ 磷酸酶处理（右）的石蜡包埋的人肺癌细胞进行免疫组织化学分析。
    Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 22 Expand Image
    使用 Fatty Acid Synthase (C20G5) Rabbit mAb 对显示脂肪细胞染色的石蜡包埋的淋巴瘤细胞进行免疫组织化学分析。
    Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 23 Expand Image
    使用 Acetyl-CoA Carboxylase (C83B10) Rabbit mAb 对石蜡包埋的人结肠癌细胞进行免疫组织化学分析。
    Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 24 Expand Image
    使用 Phospho-Acetyl-CoA Carboxylase (Ser79) (D7D11) Rabbit mAb 对未经（左）或经苯乙双胍处理（右）的石蜡包埋的 NCI-H2228 细胞团进行免疫组织化学分析。
    Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 25 Expand Image
    使用 Fatty Acid Synthase (C20G5) Rabbit mAb 对石蜡包埋的小鼠褐色脂肪进行免疫组织化学分析。
    Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 26 Expand Image
    使用 Acetyl-CoA Carboxylase (C83B10) Rabbit mAb 对石蜡包埋的人肝细胞癌细胞进行免疫组织化学分析。
    Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 27 Expand Image
    使用 Phospho-Acetyl-CoA Carboxylase (Ser79) (D7D11) Rabbit mAb（绿色）对仅营养剥夺（左上）、经血清处理（10 %，30 分钟；右上）、经 H2O2 处理（10 mM，10 分钟；左下）或经 λ 磷酸酶处理（2 小时；右下）的 293 细胞（所有细胞均在 Krebs-Ringer 碳酸氢盐缓冲液中营养剥夺 4 小时）进行共聚焦免疫荧光分析。蓝色伪彩 = DRAQ5® #4084（DNA 荧光染料）。
    Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 28 Expand Image
    使用 Acetyl-CoA Carboxylase (C83B10) Rabbit mAb 对石蜡包埋的人肺癌细胞进行免疫组织化学分析。
    Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 29 Expand Image
    使用 Fatty Acid Synthase (C20G5) Rabbit mAb（绿色）对 Hela 细胞进行共聚焦免疫荧光分析。肌动蛋白纤丝用 DY-554 Phalloidin 进行标记（红色）。蓝色伪彩 = DRAQ5™ （DNA 荧光染料）。
    Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 30 Expand Image
    对用 Acetyl-CoA Carboxylase (C83B10) Rabbit mAb（红色）标记的 NIH/3T3 细胞进行共聚焦免疫荧光分析。蓝色伪彩 = Draq5™（DNA 荧光染料）。
    Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 31 Expand Image
    使用 Acetyl-CoA Carboxylase (C83B10) Rabbit mAb（实线）或浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900（虚线）对 SK-BR-3 细胞（蓝色）和 HT-29 细胞（绿色）进行流式细胞分析。Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 用作二抗。
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    Orders: 877-616-CELL (2355)•[email protected]• Support: 877-678-TECH (8324)•[email protected]• Web: cellsignal.com
    仅供研究使用。不得用于诊断流程。