Cell Signaling Technology

Product Pathways - Screening Technologies

Phospho-AMPK Substrate Motif [LXRXX(pS/pT) MultiMab™ Rabbit mAb mix #5759

AMPK   Motif antibody   pampk   PhosphoScan   PTMScan  

No. Size Price
5759S 100 µl ( 10 western blots ) ¥3,900.00 现货查询 购买询价
5759 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 All Species Expected, Endogenous Rabbit
IP 1:100
E-P 1:1000

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, E-P=Peptide ELISA (DELFIA),

Specificity / Sensitivity

Phospho-(Ser/Thr) AMPK Substrate (P-S/T2-102) Rabbit mAb preferentially recognizes endogenous proteins and peptides bearing the LXRXXpS/pT motif. The antibody also cross-reacts with proteins and peptides that only harbor an RXXpS/pT motif.

Phospho-(Ser/Thr) AMPK Substrate (P-S/T2-102) Rabbit mAb兔单抗优先识别含有LXRXXpS/pT基序的内源性蛋白质或肽段。此抗体也可与包含RXXpS/pT基序的蛋白质和肽段发生交叉反应。

Source / Purification

Monoclonal antibody is produced by immunizing rabbits with a synthetic LXRXX(S*/T*) peptide library. The antibody is formulated from two rabbit monoclones in order to cover a broad range of reactivity.

该单克隆抗体用合成的含LXRXX(S*/T*)的肽段库免疫动物制备。该抗体是两个兔单克隆的混合,以便可以覆盖更大的反应范围。

Western Blotting

Western Blotting

Western blot analysis of extracts from H1650 cells, untreated or treated with phenformin (5 mM, 1 hr), using Phospho-(Ser/Thr) AMPK Substrate (P-S/T2-102) Rabbit mAb. Western blot was imaged using Odyssey® Infrared Imaging System (LI-COR® Biosciences).

对未处理(-)或phenformin(5 mM,1小时,+)处理的H1650细胞抽提液使用Phospho-(Ser/Thr) AMPK Substrate (P-S/T2-102) Rabbit mAb兔单抗进行Western blot分析。Western Blot图片采集使用Odyssey® Infrared Imaging System (LI-COR® Biosciences)。

Western Blotting

Western Blotting

Western blot analysis of extracts from various mouse tissues using Phospho-(Ser/Thr) AMPK Substrate (P-S/T2-102) Rabbit mAb.

对多种小鼠组织抽提液使用Phospho-(Ser/Thr) AMPK Substrate (P-S/T2-102) Rabbit mAb兔单抗进行Western blot分析。

IP

IP

Immunoprecipitation of extracts from H1650 cells using Phospho-(Ser/Thr) AMPK Substrate (P-S/T2-102) Rabbit mAb. Lane 1 shows 10% input. Western blot analysis was performed using the same antibody.

对H1650细胞抽提液使用Phospho-(Ser/Thr) AMPK Substrate (P-S/T2-102) Rabbit mAb兔单抗进行Western blot分析。列1显示10%输入对照。Western Blot分析用相同抗体进行。

Background

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 the 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, Ser108, and Ser182 (6,7). Phosphorylation at Ser108 of the β1 subunit seems to be required for the activation of AMPK enzyme, while phosphorylation at Ser24/25 and Ser182 affects AMPK localization (7). Several mutations in AMPKγ subunits have been identified, most of which are located in the putative AMP/ATP binding sites (CBS or Bateman domains). Mutations at these sites lead to reduction of AMPK activity and cause glycogen accumulation in heart or skeletal muscle (1,2). 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).

AMP-activated protein kinaseAMP激活蛋白激酶(AMPK)从酵母到动植物高度保守,在调节能量平衡上发挥关键作用(1)。AMPK是催化亚基α和调节亚基β、γ的异三聚体复合物,每个亚基由两个或三个独立基因编码(α1, 2;β1, 2;γ1, 2, 3)(2)。该激酶被细胞或环境应激时升高的AMP/ATP比率激活,如热休克、缺氧和缺血(1)。肿瘤抑制因子LKB1,与附属蛋白STRAD和MO25协同作用,磷酸化AMPKα活性环上的Thr172位点,此位点的磷酸化是AMPK活性所必需的(3-5)。AMPKα也可以在Thr258和Ser485(α1,α2为Ser491)位点上被磷酸化。这些磷酸化事件的上游激酶和生物学意义目前尚未阐清(6)。β1亚基的转录后修饰包括豆蔻酰化和多个位点的磷酸化(Ser24/25, Ser96, Ser101, Ser108, Ser182)(6,7)。β1亚基Ser108位点的磷酸化似乎是AMPK酶激活所必需,Ser24/25和Ser182位点磷酸化影响AMPK的定位(7)。AMPKγ亚基的若干突变已被发现,大部分都定位于推测的AMP/ATP结合位点(CBS或Bateman区域)。这些位点的突变导致AMPK活性降低,心脏和骨骼肌中的糖原堆积(1,2)。越来越多的证据显示AMPK不仅调节脂肪酸和糖原代谢,还通过EF2和TSC2/mTOR通路调节蛋白质合成和细胞生长,并通过eNOS/nNOS通路调节血流(1)。

AMPK phosphorylates consensus motif (L/M)XRXX(S/T)XXXL (8). Antibodies recognizing the LXRXX(S/T) motif are very useful in the identification of AMPK substrates.

AMPK磷酸化共有基序(L/M)XRXX(S/T)XXXL(8)。识别LXRXX(S/T)基序的抗体对发现AMPK底物非常有用。

  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. Lizcano, J.M. et al. (2004) EMBO J 23, 833-43.
  5. Shaw, R.J. et al. (2004) Proc Natl Acad Sci USA 101, 3329-35.
  6. Woods, A. et al. (2003) J Biol Chem 278, 28434-42.
  7. Warden, S.M. et al. (2001) Biochem J 354, 275-83.
  8. Gwinn, D.M. et al. (2008) Mol Cell 30, 214-26.

Application References

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

Use of Cell Signaling Technology (CST) Motif Antibodies within certain methods (e.g., U.S. Patents No. 7,198,896 and 7,300,753) may require a license from CST. For information regarding academic licensing terms please have your technology transfer office contact CST Legal Department at CST_ip@cellsignal.com. For information regarding commercial licensing terms please contact CST Pharma Services Department at ptmscan@cellsignal.com.

LI-COR is a registered trademark of LI-COR, Inc.

Odyssey is a registered trademark of LI-COR, Inc.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

MultiMab is a trademark of Cell Signaling Technology, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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