| 货号 | 8660T |
| 描述 | 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.PPARγ Regulated Fatty Acid Metabolism Antibody Sampler Kit 提供了一种经济的评估PPARγ 和参与脂代谢相关蛋白的方法。该试剂盒包含足够的一抗，每个可以完成四次western blots实验。 |
| 目标/特异性 | 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 (D5A2) Rabbit mAb, CBP (D6C5) Rabbit mAb, GCN5L2 (C26A10) Rabbit mAb, PPARγ (C26H12) Rabbit mAb, SirT1 (C14H4) Rabbit mAb, and RXRα (D6H10) Rabbit mAb all detect endogenous levels of their respective total proteins. |
| 供应商 | CST |
| 背景 | 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).AMPK是一个异源三聚体复合物，由α催化亚基、β和γ调控亚基组成，其中每一个都由两个或三个不同的基因（α1，2；β1，2；γ1，2，3）编码（1）。由于细胞和环境压力，如热休克、缺氧、缺血，通过升高AMP/ ATP比值激活激酶（1）。在活化环中，肿瘤抑制LKB1在苏氨酸（172位）磷酸化AMPKα，这种磷酸化对AMPK的激活是需要的（2-4）。越来越多的证据表明，AMPK不仅调节脂肪酸和糖原代谢，还通过EF2和TSC2/mTOR途径调控蛋白质合成和细胞生长，如同通过eNOS/ nNOS调节血流量（5）。CBP（CREB结合蛋白）是一种转录共激活因子，与PPARγ有关（6,7）。CBP还包含组蛋白乙酰转移酶（HAT）活性，允许其乙酰化组蛋白和其他蛋白（7）。氨基酸合成中酵母同源物2样的一般调控（GCN5L2），是一种转录接头蛋白和组蛋白乙酰转移酶（HAT）作为STAGA 和TFTC转录共激活因子复合物的催化亚基起作用（8）。GCN5L2与p300/CBP相关因子PCAF有73％的同源性，另一个HAT蛋白可在类似的复合物中找到(9)。GCN5L2乙酰化非组蛋白，如同转录共激活因子PGC1-α（10）。过氧化物酶体增殖物激活受体γ（PPARγ）是配体激活的核受体超家族成员，作为转录激活剂起作用（11）。PPARγ优先在脂肪细胞、血管平滑肌细胞和巨噬细胞表达（12）。沉默信息调节器（SIR2）家族基因是一个高度保守的基因组，编码烟酰胺腺嘌呤二核苷酸（NAD）依赖的蛋白脱乙酰基酶，也被称为III类组蛋白脱乙酰基酶（13）。SIRT1与哺乳动物的Sir2同源，是一类参与许多细胞过程调控的核蛋白，包括细胞凋亡、细胞衰老、内分泌信号、葡萄糖动态平衡、老化和长寿。 SIRT1的靶点包括PPARγ（14）和PPARγ共激活因子-1α（PGC-1α）蛋白（15）。PPARγ和PGC-1α脱乙酰化调节白色脂肪细胞在肝脏和脂肪中的糖异生/糖酵解途径的作用，响应禁食反应（14,15）。人视黄醇X受体（RXRs）由三种不同的基因（RXRα，RXRβ，RXRγ）编码，并与具有高亲和力的维生素A衍生物，9-顺-视黄酸选择性结合。核受体RXR与PPAR形成异二聚体调控脂质代谢过程（16）。Application References |
| 存放说明 | -20C |
| 参考文献 | 1 . Hardie, D.G. (2004) J Cell Sci 117, 5479-87. 2 . Goodman, R.H. and Smolik, S. (2000) Genes Dev 14, 1553-77. 3 . Guarente, L. (1999) Nat. Genet. 23, 281-285. 4 . Carling, D. (2004) Trends Biochem Sci 29, 18-24. 5 . Chan, H.M. and La Thangue, N.B. (2001) J. Cell Sci. 114, 2363-2373. 6 . Hawley, S.A. et al. (1996) J Biol Chem 271, 27879-87. 7 . Lizcano, J.M. et al. (2004) EMBO J 23, 833-43. 8 . Tontonoz, P. et al. (1995) Curr Opin Genet Dev 5, 571-6. 9 . Picard, F. et al. (2004) Nature 429, 771-776. 10 . Shaw, R.J. et al. (2004) Proc Natl Acad Sci USA 101, 3329-35. 11 . Candau, R. et al. (1996) Mol Cell Biol 16, 593-602. 12 . Rosen, E.D. et al. (1999) Mol Cell 4, 611-7. 13 . Rodgers, J.T. et al. (2005) Nature 434, 113-8. 14 . Yang, X.J. et al. (1996) Nature 382, 319-24. 15 . Lerin, C. et al. (2006) Cell Metab 3, 429-38. 16 . Gronemeyer, H. et al. (2004) Nat Rev Drug Discov 3, 950-64. |
Western blot analysis of extracts from various cell lines using GCN5L2 (C26A10) Rabbit mAb #3305.Western blot 方法检测多个细胞系提取物，使用的抗体为GCN5L2 (C26A10) Rabbit mAb #3305. | |
Western blot analysis of extracts from 293, NIH/3T3, and C6 cells using CBP (D6C5) Rabbit mAb #7389.Western blot 方法检测293细胞、NIH/3T3细胞和C6细胞提取物，使用的抗体为CBP (D6C5) Rabbit mAb #7389. | |
Western blot analysis of extracts from various cell lines using RXRα Antibody #5388.Western blot 方法检测多个细胞系提取物，使用的抗体为RXRα Antibody #5388. | |
Western blot analysis of extracts from 293, HCT15, and HeLa cells using SirT1 (C14H4) Rabbit mAb #2496.Western blot 方法检测293细胞、HCT15细胞和HeLa细胞提取物，使用的抗体为SirT1 (C14H4) Rabbit mAb #2496. | |
Western blot analysis of extracts from various cell lines and tissue using AMPKα (23A3) Rabbit mAb #2603.Western blot 方法检测多个细胞系和组织提取物，使用的抗体为AMPKα (23A3) Rabbit mAb #2603. | |
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 方法检测C2C12细胞提取物，细胞未处理或用寡霉素（0.5 µM）处理，使用的抗体为Phospho-AMPKα (Thr172) (40H9) Rabbit mAb #2535 (上图)或AMPKα Antibody #2532 (下图). | |
Western blot analysis of extracts from NIH/3T3 and 3T3-L1 cells (differentiated 6 days) using PPARγ (C26H12) Rabbit mAb #2435.Western blot 方法检测NIH/3T3 细胞和3T3-L1细胞（分化6天）提取物，使用的抗体为PPARγ (C26H12) Rabbit mAb #2435. | |
Western blot analysis of extracts from 293T cells, either mock transfected (-) or transfected with Myc/DDK-tagged cDNA expression constructs encoding full-length human RXRα (hRXRα; +), RXRβ (hRXRβ; +), or RXRγ (hRXRγ; +), using RXRα (D6H10) Rabbit mAb (upper) and DYKDDDDK Tag Antibody (Binds to same epitope as Sigmas Anti-FLAG® M2 Antibody) #2368 (lower). | |
Western blot analysis of extracts from various cell lines using RXRα (D6H10) Rabbit mAb. | |
Western blot analysis of extracts from various cell lines using CBP (D6C5) Rabbit mAb. | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 293 cells, treated with Forskolin #3828 (30 μM, 1h) and either 20 μl of CBP (D6C5) Rabbit mAb or 2 μl of Normal Rabbit IgG #2729, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using human ALS2 exon 1 primers, SimpleChIP® Human NR4A3 Promoter Primers #4829, 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. | |
Western blot analysis of extracts from HeLa, K-562, C6, and Neuro-2a cells using AMPKα (D5A2) Rabbit mAb. | |
Confocal immunofluorescent analysis of HeLa cells using CBP (D6C5) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). | |
Immunohistochemical analysis of paraffin-embedded NCI-H228 cell pellets, control (left) or phenformin-treated (right), using Phospho-AMPKalpha (T172) (40H9) Rabbit mAb. | |
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). |
