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8335
Fatty Acid and Lipid Metabolism Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Fatty Acid and Lipid Metabolism Antibody Sampler Kit #8335

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Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 1

Western blot analysis of extracts from various cell types using AceCS1 (D19C6) Rabbit mAb.

Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 2

Confocal immunofluorescent analysis of 293 cells (all nutrient-starved with Krebs-Ringer bicarbonate buffer for 4 hr), starved only (top left), serum-treated (10%, 30 min; top right), H2O2-treated (10 mM, 10 min; bottom left), or λ phosphatase-treated (2 hr; bottom right), using Phospho-Acetyl-CoA Carboxylase (Ser79) (D7D11) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 3

Flow cytometric analysis of SK-BR-3 cells (blue) and HT-29 cells (green) using Acetyl-CoA Carboxylase (C83B10) Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 4

Western blot analysis of extracts from HeLa, NIH/3T3, C6 and COS cells, using ATP-Citrate Lyase Antibody.

Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 5

Western blot analysis of extracts from NIH/3T3 cells, untreated or PDGF-treated for the indicated times, using Phospho-ATP-Citrate Lyase (Ser455) Antibody (upper) or ATP-Citrate Lyase Antibody #4332 (lower).

Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 6

Confocal immunofluorescent analysis of HeLa cells using Fatty Acid Synthase (C20G5) Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).

Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 7

Confocal immunofluorescent analysis of 3T3-L1 adipocytes (differentiated, 7 days) using Lipin 1 (D2W9G) Rabbit mAb (red). Lipid droplets were labeled with BODIPY® 493/503 (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 8

Western blot analysis of extracts from various cell lines using ACSL1 (D2H5) Rabbit mAb.

Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 9

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.

Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 10

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-Acetyl-CoA Carboxylase (Ser79) (D7D11) Rabbit mAb.

Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 11

Confocal immunofluorescent analysis of NIH/3T3 cells labeled with Acetyl-CoA Carboxylase (C83B10) Rabbit mAb (red). Blue pseudocolor=Draq5™ (fluorescent DNA dye).

Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 12

Western blot analysis of extracts from various cell lines using Lipin 1 (D2W9G) Rabbit mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).

Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 13

Immunohistochemical analysis of paraffin-embedded mouse liver using Phospho-Acetyl-CoA Carboxylase (Ser79) (D7D11) Rabbit mAb.

Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 14

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Acetyl-CoA Carboxylase (C83B10) Rabbit mAb in the presence of control peptide (left) or Acetyl-CoA Carboxylase (C83B10) Blocking Peptide #1062 (right).

Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 15

Immunohistochemical analysis of paraffin-embedded breast carcinoma using Fatty Acid Synthase (C20G5) Rabbit mAb.

Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 16

Immunohistochemical analysis of paraffin-embedded human lung carcinoma, untreated (left) or λ phosphatase-treated (right), using Phospho-Acetyl-CoA Carboxylase (Ser79) (D7D11) Rabbit mAb.

Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 17

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using Acetyl-CoA Carboxylase (C83B10) Rabbit mAb.

Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 18

Immunohistochemical analysis of paraffin-embedded hepatocellular carcinoma using Fatty Acid Synthase (C20G5) Rabbit mAb.

Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 19

Immunohistochemical analysis of paraffin-embedded NCI-H2228 cell pellets, untreated (left) or phenformin-treated (right), using Phospho-Acetyl-CoA Carboxylase (Ser79) (D7D11) Rabbit mAb.

Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 20

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Acetyl-CoA Carboxylase (C83B10) Rabbit mAb.

Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 21

Immunohistochemical analysis of paraffin-embedded lung carcinoma using Fatty Acid Synthase (C20G5) Rabbit mAb.

Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 22

Western blot analysis of extracts from SH-SY5Y cells, untreated or treated with Oligomycin #9996 (0.5 μM, 30 min), using Phospho-Acetyl-CoA Carboxylase (Ser79) (D7D11) Rabbit mAb (upper) or Acetyl-CoA Carboxylase (C83B10) Rabbit mAb #3676 (lower). The phospho-specificity of the antibody was verified by λ phosphatase treatment.

Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 23

Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma, using Acetyl-CoA Carboxylase (C83B10) Rabbit mAb.

Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 24

Immunohistochemical analysis of paraffin-embedded lymphoma, showing staining of adipocytes, using Fatty Acid Synthase (C20G5) Rabbit mAb.

Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 25

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Acetyl-CoA Carboxylase (C83B10) Rabbit mAb.

Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 26

Immunohistochemical analysis of paraffin-embedded mouse brown fat using Fatty Acid Synthase (C20G5) Rabbit mAb.

Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 27

Immunoprecipitation of Acetyl-CoA Carboxylase from HeLa cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Acetyl-CoA Carboxylase (C83B10) Rabbit mAb. Western blot was performed using Acetyl-CoA Carboxylase (C83B10) Rabbit mAb.

Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 28

Western blot analysis of extracts from various cell types using Fatty Acid Synthase (C20G5) Rabbit mAb.

Fatty Acid and Lipid Metabolism Antibody Sampler Kit: Image 29

Western blot analysis of cell extracts from various cell lines, using Acetyl-CoA Carboxylase (C83B10) Rabbit mAb.

To Purchase # 8335T
Product # Size Price
8335T
1 Kit  (8 x 20 µl) $ 554

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
AceCS1 (D19C6) Rabbit mAb 3658 20 µl
  • WB
  • IP
H M R Mk 78 Rabbit IgG
Phospho-Acetyl-CoA Carboxylase (Ser79) (D7D11) Rabbit mAb 11818 20 µl
  • WB
  • IP
  • IHC
  • IF
H M R 280 Rabbit IgG
Acetyl-CoA Carboxylase (C83B10) Rabbit mAb 3676 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
H M R Hm 280 Rabbit IgG
ATP-Citrate Lyase Antibody 4332 20 µl
  • WB
H M R Mk 125 Rabbit 
Phospho-ATP-Citrate Lyase (Ser455) Antibody 4331 20 µl
  • WB
  • IP
H M 125 Rabbit 
Fatty Acid Synthase (C20G5) Rabbit mAb 3180 20 µl
  • WB
  • IP
  • IHC
  • IF
H M R 273 Rabbit IgG
Lipin 1 (D2W9G) Rabbit mAb 14906 20 µl
  • WB
  • IP
  • IF
H M 130 Rabbit IgG
ACSL1 (D2H5) Rabbit mAb 9189 20 µl
  • WB
  • IP
H M R Mk 78 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

Product 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.

Specificity / Sensitivity

AceCS1 (D19C6) Rabbit mAb recognizes endogenous levels of total cytoplasmic acetyl-CoA synthetase. Phospho-Acetyl-CoA Carboxylase (Ser79) (D7D11) Rabbit mAb recognizes endogenous levels of ACC only when phosphorylated at Ser79; this antibody recognizes both ACCα and ACCß. Acetyl-CoA Carboxylase (C83B10) Rabbit mAb recognizes endogenous levels of all isoforms of acetyl-CoA carboxylase protein. Phospho-ATP-Citrate Lyase (Ser455) Antibody recognizes endogenous levels of ATP-citrate lyase only when phosphorylated at Ser455. ACSL1 (D2H5) Rabbit mAb, ATP-Citrate Lyase Antibody, Fatty Acid Synthase (C20G5) Rabbit mAb, and Lipin 1 (D2W9G) Rabbit mAb recognize endogenous levels of their respective target proteins.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the sequence of human cytoplasmic acetyl-CoA synthetase, residues surrounding Ser523 of human acetyl-CoA carboxylase α1, residues surrounding Gly46 of human fatty acid synthase, residues near the carboxy terminus of human lipin 1 or residues surrounding Ala257 of human ACSL1 protein. Monoclonal activation state antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser79 of human acetyl-CoA carboxylase protein. Modification state-specific polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser455 of human ATP-citrate lyase protein. Polyclonal antibodies are produced by immunizing animals corresponding to residues of human ATP-citrate lyase protein. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.

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.

Limited Uses

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