| 货号 | 9957T |
| 描述 | The AMPK and ACC Antibody Sampler Kit provides an economical means to investigate energy homeostasis and fatty acid synthesis within the cell. The kit contains primary and secondary antibodies to perform four Western blots with each antibody. AMPK和ACC抗体试剂盒为细胞内的能量平衡和脂肪酸合成的研究提供了一种经济有效的方法。试剂盒同时提供一抗和二抗，每个抗体可以做四次Western blot. |
| 目标/特异性 | Phospho-AMPKα (Thr172) (40H9) Rabbit mAb detects endogenous AMPKα only when phosphorylated at Thr172. The antibody detects both α1 and α2 isoforms of the catalytic subunit, but does not detect the regulatory β or γ subunits. AMPKα (D5A2) Rabbit mAb detects endogenous levels of total AMPKα protein. Phospho-AMPKβ1 (Ser182) Antibody detects endogenous levels of AMPKβ1 only when phosphorylated at Ser182. AMPKβ1/2 (57C12) Rabbit mAb detects endogenous levels of both total AMPKβ1 and β2 proteins. The antibody does not cross-react with other related proteins. Phospho-Acetyl-CoA Carboxylase (Ser79) (D7D11) Rabbit mAb recognizes endogenous levels of acetyl-CoA carboxylase protein only when phosphorylated at Ser79. Acetyl-CoA Carboxylase (C83B10) Rabbit mAb detects endogenous levels of all isoforms of acetyl-CoA carboxylase protein. |
| 供应商 | CST |
| 背景 | AMP-activated protein kinase (AMPK) is highly conserved from yeast to plants and animals and plays a key role in the regulation of energy homeostasis (1). 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)(2). 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, in association with accessory proteins STRAD and MO25, phosphorylates AMPKα at Thr172 in the activation loop and this phosphorylation is required for AMPK activation (3-5). AMPKα is also phosphorylated at Thr258 and Ser485 (for α1; Ser491 for α2). The upstream kinase and biological significance of these phosphorylation events have yet to be elucidated (6). The β1 subunit is post-translationally modified by myristoylation and multi-site phosphorylation including Ser24/25, Ser96, Ser101 and Ser182 (6,7). Phosphorylation at Ser108 of the β1 subunit seems to be required for the activation of AMPK enzyme, while phosphorylation ot Ser24/25 and Ser182 affects AMPK localization (7). 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 (1).
Acetyl-CoA carboxylase (ACC) catalyzes the pivotal step of the fatty acid synthesis pathway. The 265 kDa ACCα is the predominant isoform found in liver, adipocytes and mammary gland, while the 280 kDa ACCβ is the major isoform in skeletal muscle and heart (8). Phosphorylation by AMPK at Ser79 or by PKA at Ser1200 inhibits the enzymatic activity of ACC (9). ACC is a potential target of anti-obesity drugs (10,11). 腺苷酸激活的蛋白激酶（AMPK）是一种在酵母，植物和动物界高度保守的蛋白激酶，它在能量平衡中发挥关键性的作用(1)。AMPK是异源三聚体复合物，由α催化亚基和β、γ两个调节亚基组成，每个亚基都由两到三个不同的基因编码(α1, 2; β1, 2; γ1, 2, 3)(2)。细胞或者环境因素的刺激，如热激，缺氧，缺血等，导致AMP/ATP浓度比的升高，AMP/ATP浓度比的升高可以激活AMPK。肿瘤抑制因子LKB1，与辅助蛋白STRAD和MO25相结合，在AMPK的α亚基活化环的172位苏氨酸上对其磷酸化，磷酸化对AMPK的激活是必需的(3-5)。AMPKα同样也可以在Thr258和Ser485(α1; α2是Ser491)位点磷酸化。此过程的上游激酶和这些位点磷酸化的生物学意义尚不明晰(6)。β1亚基有一些翻译后的修饰，包括豆蒄酰化修饰，以及在多个位点的磷酸化修饰(包括Ser24/25, Ser96, Ser101和Ser182)(6,7)。β1亚基Ser108的磷酸化可能对AMPK的激活来说是必需的，而Ser24/25和Ser182的磷酸化可能影响到AMPK的定位(7)。越来越多的证据表明，AMPK不仅调控机体的糖脂代谢，同时也通过EF2和TSC2/mTOR通路调控细胞生长和蛋白质的合成，以及通过eNOS/nNOS系统来调节血流量(1)。乙酰辅酶A羧化酶(ACC)在脂肪酸合成的关键步骤中起着催化作用。分子量为265 kDa的ACCα，是肝脏，脂肪细胞和乳腺中ACC的主要形式，而在骨胳肌和心脏中，其主要形式为280 kDa的ACCβ(8)。ACC可以在Ser79被AMPK磷酸化，或者在Ser1200被PKA磷酸化，这些位点的磷酸化可以抑制ACC的酶活性(9)。ACC也是减肥药物的一个潜在靶点(10,11)。Application References |
| 存放说明 | -20C |
| 参考文献 | 1 . Hardie, D.G. (2004) J Cell Sci 117, 5479-87. 2 . Carling, D. (2004) Trends Biochem Sci 29, 18-24. 3 . Hawley, S.A. et al. (1996) J Biol Chem 271, 27879-87. 4 . Ha, J. et al. (1994) J Biol Chem 269, 22162-8. 5 . Lizcano, J.M. et al. (2004) EMBO J 23, 833-43. 6 . Abu-Elheiga, L. et al. (2001) Science 291, 2613-6. 7 . Levert, K.L. et al. (2002) J Biol Chem 277, 16347-50. 8 . Woods, A. et al. (2003) J Biol Chem 278, 28434-42. 9 . Warden, S.M. et al. (2001) Biochem J 354, 275-83. 10 . Shaw, R.J. et al. (2004) Proc Natl Acad Sci USA 101, 3329-35. 11 . Ruderman, N.B. et al. (1999) Am. J. Physiol. 276, E1-E18. |
Immunohistochemical analysis of paraffin-embedded mouse liver using Phospho-Acetyl-CoA Carboxylase (Ser79) (D7D11) Rabbit mAb. | |
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-Acetyl-CoA Carboxylase (Ser79) (D7D11) Rabbit mAb. | |
Immunohistochemical analysis of paraffin-embedded human lung carcinoma, untreated (left) or λ phosphatase-treated (right), using Phospho-Acetyl-CoA Carboxylase (Ser79) (D7D11) Rabbit mAb. | |
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. | |
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). | |
Immunohistochemical analysis of paraffin-embedded NCI-H2228 cell pellets, untreated (left) or phenformin-treated (right), using Phospho-Acetyl-CoA Carboxylase (Ser79) (D7D11) Rabbit mAb. | |
Immunohistochemical analysis of paraffin-embedded NCI-H228 cell pellets, control (left) or phenformin-treated (right), using Phospho-AMPKalpha (T172) (40H9) Rabbit mAb. | |
Western blot analysis of extracts from HeLa, K-562, C6, and Neuro-2a cells using AMPKα (D5A2) Rabbit mAb. | |
Western blot analysis of extracts from various cell types using Acetyl-CoA Carboxylase (C83B10) Rabbit mAb #3676. | |
Western blot analysis of cell lysates from various cell types using AMPKβ1/2 (57C12) Rabbit mAb #4150. | |
Western blot analysis of extracts from C2C12 cells, untreated (lanes 1,3) or oligomycin-treated (lanes 2,4), using Phospho-AMPKβ1 (Ser108) Antibody #4181 (upper) or AMPKβ1 Antibody #4182 (lower). Cell lysates were treated with λ phosphatase in lanes 3 and 4 to demonstrate the phospho-specificity of Phospho-AMPKβ1 (Ser108) Antibody. | |
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 HEK293 cells, untreated or oligomycin-treated, using Phospho-Acetyl-CoA Carboxylase (Ser79) Antibody #3661. | |
Western blot analysis of extracts from various cells and tissues using AMPKα (23A3) Rabbit mAb #2603. | |
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). |
