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12589
Adipogenesis Marker Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit
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Adipogenesis Marker Antibody Sampler Kit #12589

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Adipogenesis Marker Antibody Sampler Kit: Image 1
Western blot analysis of extracts from NIH/3T3 and 3T3-L1 cells (differentiated 6 days into adipocytes) using PPARγ (C26H12) Rabbit mAb.
Adipogenesis Marker Antibody Sampler Kit: Image 2
Immunohistochemical analysis of 3T3-L1 cells, undifferentiated (left) or differentiated (right) , using PPARγ (C26H12) Rabbit mAb.
Adipogenesis Marker Antibody Sampler Kit: Image 3
Immunohistochemical analysis of paraffin-embedded mouse brown fat using PPARγ (C26H12) Rabbit mAb.
Adipogenesis Marker Antibody Sampler Kit: Image 4
Confocal immunofluorescent analysis of 3T3-L1 cells using PPARγ (C26H12A8) Rabbit mAb (red) showing nuclear localization in differentiated cells. Lipid droplets have been labeled with BODIPY 493/503 (green). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).
Adipogenesis Marker Antibody Sampler Kit: Image 5
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.
Adipogenesis Marker Antibody Sampler Kit: Image 6
Western blot analysis of extracts from NIH/3T3 and 3T3-L1 cells using FABP4 Antibody.
Adipogenesis Marker Antibody Sampler Kit: Image 7
Chromatin immunoprecipitations were performed with cross-linked chromatin from untreated HDLM-2 cells and PPARγ (C26H12) Rabbit mAb using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Library was prepared using DNA Library Prep Kit for Illumina (ChIP-seq, CUT&RUN) #56795. The figure shows binding across FOXN3, a known target gene of PPARγ.
Adipogenesis Marker Antibody Sampler Kit: Image 8
Western blot analysis of extracts from NIH/3T3 and 3T3-L1 cells (differentiated into adipocytes) using Adiponectin (C45B10) Rabbit mAb.
Adipogenesis Marker Antibody Sampler Kit: Image 9
Western blot analysis of extracts from various cell types using Fatty Acid Synthase (C20G5) Rabbit mAb.
Adipogenesis Marker Antibody Sampler Kit: Image 10
Western blot analysis of cell extracts from various cell lines, using Acetyl-CoA Carboxylase (C83B10) Rabbit mAb.
Adipogenesis Marker Antibody Sampler Kit: Image 11
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.
Adipogenesis Marker Antibody Sampler Kit: Image 12
Western blot analysis of extracts from Hep G2 and LNCaP cells using C/EBPα (D56F10) XP® Rabbit mAb.
Adipogenesis Marker Antibody Sampler Kit: Image 13
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).
Adipogenesis Marker Antibody Sampler Kit: Image 14
Chromatin immunoprecipitations were performed with cross-linked chromatin from untreated HDLM-2 cells and PPARγ (C26H12) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Library was prepared using DNA Library Prep Kit for Illumina (ChIP-seq, CUT&RUN) #56795. The figures show binding across chromosome 14 (upper), including FOXN3 (lower), a known target gene of PPARγ.
Adipogenesis Marker Antibody Sampler Kit: Image 15
Immunohistochemical analysis of paraffin-embedded breast carcinoma using Fatty Acid Synthase (C20G5) Rabbit mAb.
Adipogenesis Marker Antibody Sampler Kit: Image 16
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.
Adipogenesis Marker Antibody Sampler Kit: Image 17
Immunohistochemical analysis of paraffin-embedded mouse lung using C/EBPα (D56F10) XP® Rabbit mAb.
Adipogenesis Marker Antibody Sampler Kit: Image 18
Immunohistochemical analysis of paraffin-embedded breast carcinoma using Perilipin-1 (D1D8) XP® Rabbit mAb. Note specific staining of adipocytes.
Adipogenesis Marker Antibody Sampler Kit: Image 19
Chromatin immunoprecipitations were performed with cross-linked chromatin from HDLM-2 cells and either PPARγ (C26H12) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human FOXN3 Intron 3 Primers #95568, human STON2 intron 4 primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
Adipogenesis Marker Antibody Sampler Kit: Image 20
Immunohistochemical analysis of paraffin-embedded hepatocellular carcinoma using Fatty Acid Synthase (C20G5) Rabbit mAb.
Adipogenesis Marker Antibody Sampler Kit: Image 21
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).
Adipogenesis Marker Antibody Sampler Kit: Image 22
Immunohistochemical analysis of paraffin-embedded cell pellets, THP-1 (left) or Jurkat (right), using C/EBPα (D56F10) XP® Rabbit mAb.
Adipogenesis Marker Antibody Sampler Kit: Image 23
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).
Adipogenesis Marker Antibody Sampler Kit: Image 24
Immunohistochemical analysis of paraffin-embedded lung carcinoma using Fatty Acid Synthase (C20G5) Rabbit mAb.
Adipogenesis Marker Antibody Sampler Kit: Image 25
Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using Acetyl-CoA Carboxylase (C83B10) Rabbit mAb.
Adipogenesis Marker Antibody Sampler Kit: Image 26
Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma using C/EBPα (D56F10) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
Adipogenesis Marker Antibody Sampler Kit: Image 27
Immunohistochemical analysis of parafin-embedded mouse kidney with staining of adjacent adipose tissue using Perilipin-1 (D1D8) XP® Rabbit mAb.
Adipogenesis Marker Antibody Sampler Kit: Image 28
Immunohistochemical analysis of paraffin-embedded lymphoma, showing staining of adipocytes, using Fatty Acid Synthase (C20G5) Rabbit mAb.
Adipogenesis Marker Antibody Sampler Kit: Image 29
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Acetyl-CoA Carboxylase (C83B10) Rabbit mAb.
Adipogenesis Marker Antibody Sampler Kit: Image 30
Immunohistochemical analysis of paraffin-embedded human tonsil using C/EBPα (D56F10) XP® Rabbit mAb.
Adipogenesis Marker Antibody Sampler Kit: Image 31
Immunohistochemical analysis of parafin-embedded mouse prostate with staining of adjacent adipose tissue using Perilipin-1 (D1D8) XP® Rabbit mAb.
Adipogenesis Marker Antibody Sampler Kit: Image 32
Immunohistochemical analysis of paraffin-embedded mouse brown fat using Fatty Acid Synthase (C20G5) Rabbit mAb.
Adipogenesis Marker Antibody Sampler Kit: Image 33
Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma, using Acetyl-CoA Carboxylase (C83B10) Rabbit mAb.
Adipogenesis Marker Antibody Sampler Kit: Image 34
Confocal immunofluorescent analysis of THP-1 (left) and Jurkat (right) cells using C/EBPα (D56F10) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Adipogenesis Marker Antibody Sampler Kit: Image 35
Immunohistochemical analysis of parafin-embedded mouse testis with staining of adjacent adipose tissue using Perilipin-1 (D1D8) XP® Rabbit mAb.
Adipogenesis Marker Antibody Sampler Kit: Image 36
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Acetyl-CoA Carboxylase (C83B10) Rabbit mAb.
Adipogenesis Marker Antibody Sampler Kit: Image 37
Confocal immunofluorescent analysis of differentiated 3T3-L1 cells using C/EBPα (D56F10) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Adipogenesis Marker Antibody Sampler Kit: Image 38
Confocal immunofluorescent analysis of frozen mouse brown adipose tissue using Perilipin-1 (D1D8) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Adipogenesis Marker Antibody Sampler Kit: Image 39
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).
Adipogenesis Marker Antibody Sampler Kit: Image 40
Confocal immunofluorescent analysis of NIH/3T3 cells labeled with Acetyl-CoA Carboxylase (C83B10) Rabbit mAb (red). Blue pseudocolor=Draq5™ (fluorescent DNA dye).
Adipogenesis Marker Antibody Sampler Kit: Image 41
Confocal immunofluorescent analysis of 8-day differentiated 3T3-L1 cells, using Perilipin (D1D8) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Adipogenesis Marker Antibody Sampler Kit: Image 42
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.
Adipogenesis Marker Antibody Sampler Kit: Image 43
Flow cytometric analysis of THP-1 cells using C/EBP-α (D56F10) XP® Rabbit mAb (blue) compared to concentration-mathced Rabbit (DA1E) mAb IgG XP® Isotype Contol #3900 (red). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody
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To Purchase # 12589
製品# サイズ 数量 価格 在庫
12589T		 1 Kit  (7 x 20 microliters)

Custom Formulation

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Acetyl-CoA Carboxylase (C83B10) Rabbit mAb 3676 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
 H M R Hm 280 Rabbit IgG
Adiponectin (C45B10) Rabbit mAb 2789 20 µl
  • WB
 H M R 27 Rabbit IgG
C/EBPα (D56F10) XP® Rabbit mAb 8178 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
 H M 42, 28 Rabbit IgG
FABP4 Antibody 2120 20 µl
  • WB
 H M 15 Rabbit 
Fatty Acid Synthase (C20G5) Rabbit mAb 3180 20 µl
  • WB
  • IP
  • IHC
  • IF
 H M R 273 Rabbit IgG
Perilipin-1 (D1D8) XP® Rabbit mAb 9349 20 µl
  • WB
  • IP
  • IHC
  • IF
 H M 62 Rabbit IgG
PPARγ (C26H12) Rabbit mAb 2435 20 µl
  • WB
  • IHC
  • IF
  • ChIP
 H M 53, 57 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
 Rab Goat 

Product Description

The Adipogenesis Marker Antibody Sampler Kit provides an economical means to evaluate proteins involved in the regulation of adipogenesis. The kit includes enough antibody to perform two western blot experiments with each primary antibody.

Specificity / Sensitivity

Each antibody recognizes endogenous total levels of its specific target protein. The Adiponectin (C45B10) Rabbit mAb detects endogenous levels of total adiponectin protein monomer. It will not detect higher molecular weight forms of adiponectin. The Acetyl-CoA Carboxylase (C83B10) Rabbit mAb detects endogenous levels of all isoforms of acetyl-CoA carboxylase protein. The FABP4 Antibody may cross react with other FABP family members.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser523 of human acetyl-CoA carboxylase α1, to human adiponectin, to the sequence of mouse FABP4, to residues surrounding Gly46 of human fatty acid synthase, to residues surrounding Ile419 of human perilipin/perilipin-1 protein, to residues surrounding Ala176 of human C/EBPα protein, or to residues surrounding Asp69 of human PPARγ.

Background

Adipocytes are the primary cellular component of adipose tissue and play a key role in the storage of triacylglycerol. Adipogenesis is the cellular process where preadipocytes differentiate into adipocytes.

Fatty acid binding proteins (FABPs) act as cytoplasmic lipid chaperones by binding fatty acids and lipids for transport to various cellular pathways (1,2). The predominant fatty acid binding protein found in adipocytes is FABP4.

Adiponectin is an adipokine expressed exclusively in brown and white adipocytes and is secreted into the blood. It exists in three major forms: a low molecular weight trimer, a medium molecular weight hexamer and a high molecular weight multimer (3). Decreased adiponectin levels are seen in obese and insulin-resistant mice and humans (4), suggesting that this adipokine is critical for maintenance of insulin sensitivity.

Peroxisome proliferator-activated receptor γ (PPARγ) is a transcriptional activator preferentially expressed in adipocytes, vascular smooth muscle cells, and macrophages (5,6).

Acetyl-CoA carboxylase (ACC) is a key fatty acid biosynthesis and oxidation enzyme that is responsible for the carboxylation of acetyl-CoA to malonyl-CoA, (7). Phosphorylation of acetyl-CoA carboxylase by AMPK at Ser79 or by PKA at Ser1200 inhibits ACC enzymatic activity (8). ACC is a potential target of anti-obesity drugs (9,10).

CCAAT/enhancer-binding proteins (C/EBPs) transcription factors are critical for cellular differentiation, terminal function, and the inflammatory response (11). Phosphorylation of C/EBPα at Thr222, Thr226, and Ser230 by GSK-3 may be required for adipogenesis (12).

Perilipin localizes to the periphery of lipid droplets and serves as a protective coating against lipases. Evidence suggests that PKA regulates lipolysis by phosphorylating perilipin (13-17), resulting in a conformational change that exposes lipid droplets to endogenous, hormone-sensitive lipases (14). Hence, perilipin plays a pivotal role in lipid storage (14,17).

Fatty acid synthase (FASN) catalyzes the synthesis of long-chain fatty acids from acetyl-CoA and malonyl-CoA. FASN is active as a homodimer with seven different catalytic activities and produces lipids in the liver for export to metabolically active tissues or storage in adipose tissue. In most other human tissues, FASN is minimally expressed since they rely on circulating fatty acids for new structural lipid synthesis (18).
  1. Tuncman, G. et al. (2006) Proc Natl Acad Sci U S A 103, 6970-5.
  2. Haunerland, N.H. and Spener, F. (2004) Prog Lipid Res 43, 328-49.
  3. Kadowaki, T. et al. (2006) J Clin Invest 116, 1784-92.
  4. Hu, E. et al. (1996) J Biol Chem 271, 10697-703.
  5. Tontonoz, P. et al. (1995) Curr Opin Genet Dev 5, 571-6.
  6. Rosen, E.D. et al. (1999) Mol Cell 4, 611-7.
  7. Castle, J.C. et al. (2009) PLoS One 4, e4369.
  8. Ha, J. et al. (1994) J Biol Chem 269, 22162-8.
  9. Abu-Elheiga, L. et al. (2001) Science 291, 2613-6.
  10. Levert, K.L. et al. (2002) J Biol Chem 277, 16347-50.
  11. Lekstrom-Himes, J. and Xanthopoulos, K.G. (1998) J Biol Chem 273, 28545-8.
  12. Ross, S.E. et al. (1999) Mol Cell Biol 19, 8433-41.
  13. Greenberg, A.S. et al. (1991) J Biol Chem 266, 11341-6.
  14. Brasaemle, D.L. (2007) J Lipid Res 48, 2547-59.
  15. Ducharme, N.A. and Bickel, P.E. (2008) Endocrinology 149, 942-9.
  16. Egan, J.J. et al. (1990) J Biol Chem 265, 18769-75.
  17. Brasaemle, D.L. et al. (2009) Mol Cell Biochem 326, 15-21.
  18. Katsurada, A. et al. (1990) Eur J Biochem 190, 427-33.

Pathways & Proteins

Explore pathways + proteins related to this product.

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U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
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