Product Pathways - Nuclear Receptor Signaling
PPARγ Regulated Fatty Acid Metabolism Antibody Sampler Kit #8660
|Kit Includes||Quantity||Applications||Reactivity||MW (kDa)||Isotype|
|Phospho-AMPKα (Thr172) (40H9) Rabbit mAb #2535||40 µl||W IP IHC-P||H M R Hm Mk Dm Sc (C) (Z) (B) (Pg)||62||Rabbit IgG|
|AMPKα (23A3) Rabbit mAb #2603||40 µl||W||H M R Mk||62||Rabbit IgG|
|CBP (D6C5) Rabbit mAb #7389||40 µl||W IP IF-IC ChIP||H M R Mk||300||Rabbit IgG|
|GCN5L2 (C26A10) Rabbit mAb #3305||40 µl||W IP IF-IC||H M R Mk (B) (Hr)||94||Rabbit IgG|
|PPARγ (C26H12) Rabbit mAb #2435||40 µl||W IHC-P IF-IC||H M (R)||53, 57||Rabbit IgG|
|SirT1 (C14H4) Rabbit mAb #2496||40 µl||W IP||H||120||Rabbit|
|RXRα Antibody #5388||40 µl||W IP||H R Mk||53||Rabbit|
|Anti-rabbit IgG, HRP-linked Antibody #7074||100 µl||Goat|
Reactivity Key: H=Human M=Mouse R=Rat Hm=Hamster Mk=Monkey C=Chicken Dm=D. melanogaster Z=Zebrafish B=Bovine Pg=Pig Sc=S. cerevisiae Hr=Horse
Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Specificity / Sensitivity
Phospho-AMPKα (Thr172) (40H9) Rabbit mAb detects endogenous AMPKα only when phosphorylated at Thr172. Phospho-AMPKα (Thr172) (40H9) Rabbit mAb detects both α1 and α2 isoforms of the catalytic subunit, but does not detect the regulatory β or γ subunits. AMPKα (23A3) Rabbit mAb, CBP (D6C5) Rabbit mAb, GCN5L2 (C26A10) Rabbit mAb, PPARγ (C26H12) Rabbit mAb, SirT1 (C14H4) Rabbit mAb, and RXRα Antibody all detect endogenous levels of their respective total proteins.
Western blot analysis of extracts from NIH/3T3 and 3T3-L1 cells (differentiated 6 days) using PPARγ (C26H12) Rabbit mAb #2435.
Western blot analysis of extracts from 293, HCT15, and HeLa cells using SirT1 (C14H4) Rabbit mAb #2496.
Western blot analysis of extracts from C2C12 cells, untreated or oligomycin-treated (0.5 µM), using Phospho-AMPKα (Thr172) (40H9) Rabbit mAb #2535 (upper) or AMPKα Antibody #2532 (lower).
Western blot analysis of extracts from various cell lines and tissue using AMPKα (23A3) Rabbit mAb #2603.
Western blot analysis of extracts from various cell lines using GCN5L2 (C26A10) Rabbit mAb #3305.
Western blot analysis of extracts from various cell lines using RXRα Antibody #5388.
PPARγ Regulated Fatty Acid Metabolism Antibody Sampler Kit provides an economical means to evaluate PPARγ and related proteins involved in lipid metabolism. This kit contains enough primary antibody to perform four western blots per primary.
Source / Purification
Monoclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr172 of human AMPKα protein or with a synthetic peptide corresponding to the respective sequences of human AMPKα, CBP, GCN5L2, PPARγ, and SirT1. Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human RXRα protein. Antibodies are purified by protein A and peptide affinity chromatography.
AMPK is a heterotrimeric complex composed of a catalytic α subunit and regulatory β and γ subunits, each of which is encoded by two or three distinct genes (α1, 2; β1, 2; γ1, 2, 3) (1). The kinase is activated by an elevated AMP/ATP ratio due to cellular and environmental stress, such as heat shock, hypoxia, and ischemia (1). The tumor suppressor LKB1 phosphorylates AMPKα at Thr172 in the activation loop, and this phosphorylation is required for AMPK activation (2-4). Accumulating evidence indicates that AMPK not only regulates the metabolism of fatty acids and glycogen, but also modulates protein synthesis and cell growth through EF2 and TSC2/mTOR pathways, as well as blood flow via eNOS/nNOS (5).CBP (CREB-binding protein) is a transcriptional co-activator that associates with PPARγ (6,7). CBP also contains histone acetyltransferase (HAT) activity, allowing it to acetylate histones and other proteins (7).General Control of Amino Acid Synthesis Yeast Homolog Like 2 (GCN5L2) is a transcription adaptor protein and a histone acetyltransferase (HAT) that functions as the catalytic subunit of the STAGA and TFTC transcription coactivator complexes (8). GCN5L2 is 73% homologous to the p300/CBP-associated factor PCAF, another HAT protein found in similar complexes (9). GCN5L2 acetylates non-histone proteins such as the transcription co-activator PGC1-α (10).Peroxisome proliferator-activated receptor γ (PPARγ) is a member of the ligand-activated nuclear receptor superfamily and functions as a transcriptional activator (11). PPARγ is preferentially expressed in adipocytes as well as in vascular smooth muscle cells and macrophage (12).The Silent Information Regulator (SIR2) family of genes is a highly conserved group of genes that encode nicotinamide adenine dinucleotide (NAD)-dependent protein deacetylases, also known as class III histone deacetylases (13). SirT1, the mammalian ortholog of Sir2, is a nuclear protein implicated in the regulation of many cellular processes, including apoptosis, cellular senescence, endocrine signaling, glucose homeostasis, aging, and longevity. Targets of SirT1 include PPARγ (14), and the PPARγ coactivator-1α (PGC-1α) protein (15). Deacetylation of PPARγ and PGC-1α regulates the gluconeogenic/glycolytic pathways in the liver and fat mobilization in white adipocytes in response to fasting (14,15).The human retinoid X receptors (RXRs) are type-II nuclear hormone receptors encoded by three distinct genes (RXRα, RXRβ, and RXRγ) and bind selectively and with high affinity to the vitamin A derivative, 9-cis-retinoic acid. Nuclear RXRs form heterodimers with PPAR to help regulate transcription during lipid metabolism (16).
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- Chan, H.M. and La Thangue, N.B. (2001) J Cell Sci 114, 2363-73.
- Candau, R. et al. (1996) Mol Cell Biol 16, 593-602.
- Yang, X.J. et al. (1996) Nature 382, 319-24.
- Lerin, C. et al. (2006) Cell Metab 3, 429-38.
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Selected rabbit monocloal antibodies are produced under license (granting certain rights including those under U. S. Patents No. 5,675,063 and in some instances 7,429,487) from Epitomics, Inc.
For Research Use Only. Not For Use In Diagnostic Procedures.