Cell Signaling Technology

Product Pathways - PI3K / Akt Signaling

Phospho-NDRG1 (Thr346) (D98G11) XP® Rabbit mAb #5482

ndrg   ndrg1  

No. Size Price
5482S 100 µl ( 10 western blots ) ¥4,050.00 现货查询 购买询价
5482T 20 µl ( 2 western blots ) ¥1,600.00 现货查询 购买询价
5482 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat,Monkey, Endogenous 46, 48 Rabbit IgG
IHC-P 1:500
F 1:200
IF-IC 1:800

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IHC-P=Immunohistochemistry (Paraffin), F=Flow Cytometry, IF-IC=Immunofluorescence (Immunocytochemistry),

Specificity / Sensitivity

Phospho-NDRG1 (Thr346) (D98G11) XP® Rabbit mAb (HRP Conjugate) detects endogenous levels of NDRG1 when phosphorylated at Thr346. This antibody likely cross-reacts with other conserved phosporylation sites on NDRG1 at positions Thr356 and Thr366.

Phospho-NDRG1 (Thr346) (D98G11) XP® Rabbit mAb可以识别Thr346被磷酸化的内源性NDRG1蛋白。另外此抗体和Thr356及Thr366被磷酸化的NDRG1蛋白貌似有交叉反应。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr346 of NDRG1.

单克隆抗体是通过一种合成的磷酸化肽段去免疫动物产生。这种合成的肽段与NDRG1邻近Thr346 氨基酸残基序列一致。

IF-IC

IF-IC

Confocal immunofluorescent analysis of C2C12 cells, treated with LY294002 #9901 (left) or insulin (right), using Phospho-NDRG1 (Thr346) (D98G11) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

使用LY294002#9901(左)或胰岛素(右)处理C2C12细胞,使用Phospho-NDRG1 (Thr346) (D98G11) XP® 兔单抗(绿色)进行激光共聚焦荧光分析。用DY-554phalloidin(红色)标记肌蛋白丝。蓝色伪彩=DRAQ5® #4084 (荧光染料)。

Western Blotting

Western Blotting

Western blot analysis of extracts from C2C12 cells, untreated or treated with LY294002 #9901, insulin, or both using Phospho-NDRG1 (Thr346) (D98G11) XP® Rabbit mAb (upper) or total NDRG1 Antibody #5196 (lower).

未经LY294002 #9901, insulin或经两者共同处理后的C2C12细胞获得提取物进行western blot实验,使用Phospho-NDRG1 (Thr346) (D98G11) XP® 兔单抗(上) 或total NDRG1 抗体 #5196 (下).

Western Blotting

Western Blotting

Western blot analysis of extracts from A431 cells treated with hEGF #8916 (100 ng/ml, 30 minutes), with or without calf intestinal phosphatase and λ-phosphatase, using Phospho-NDRG1 (Thr346) (D98G11) XP® Rabbit mAb (upper) or total NDRG1 Antibody #5196 (lower).

在牛小肠碱性磷酸酶和λ-磷酸酶存在或不存在的条件下使用hEGF#8916(100ng/ml,30分钟)处理A431细胞,获得提取物进行western blot实验,使用Phospho-NDRG1 (Thr346) (D98G11) XP® 兔单抗(上) 或total NDRG1 抗体#5196 (下)检测。

Western Blotting

Western Blotting

Western blot analysis of HeLa cells treated with hEGF #8916 (100 ng/ml, 30 minutes), mock transfected or transfected with SignalSilence® NDRG1 siRNA I #6245 or SignalSilence® NDRG1 siRNA II #6257, using Phospho-NDRG1 (Thr346) (D98G11) XP® Rabbit mAb (upper) or β-Tubulin (9F3) Rabbit mAb #2128 (lower).

HeLa细胞经hEGF #8916 (100 ng/ml, 30 分钟),未转染或转染SignalSilence® NDRG1 siRNA I #6245 或SignalSilence® NDRG1 siRNA II #6257后获得细胞提取物进行western blot实验,使用using Phospho-NDRG1 (Thr346) (D98G11) XP® 兔单抗 (上)或 β-Tubulin (9F3)兔单抗 #2128 (下)进行检测。

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells, untreated (green) or treated with LY294002 #9901, wortmannin #9951 and U0126 #9903 (blue), using Phospho-NDRG1 (Thr346) (D98G11) XP® Rabbit mAb.

未处理(绿色)或经LY294002#9901,wortmannin#9951和U0126 #9903(蓝色)处理后的Jurkat细胞,使用Phospho-NDRG1 (Thr346) (D98G11) XP® 兔单抗进行流式细胞分析。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-emebedded human lung carcinoma using Phospho-NDRG1 (Thr346) (D98G11) XP® Rabbit mAb.

使用Phospho-NDRG1 (Thr346) (D98G11) XP® 兔单抗对石蜡包埋的人肺癌组织进行免疫组化分析。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis on SignalSlide® Phospho-Akt (Ser473) IHC Controls #8101 [paraffin-embedded LNCaP cell pellets, control (left) or LY294002-treated (right)] using NDRG1 Antibody #5196 (top) or Phospho-NDRG1 (Thr346) (D98G11) XP® Rabbit mAb (bottom).

SignalSlide® Phospho-Akt (Ser473) IHC对照#8101[石蜡包埋 LNCaP 细胞, 对照 (左) 或 LY294002-处理(右)] ,使用NDRG1 抗体#5196(上)或Phospho-NDRG1 (Thr346) (D98G11) XP® 兔单抗(下)进行免疫组化分析。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human cervical carcinoma, control (left) or λ phosphatase-treated (right), using Phospho-NDRG1 (Thr346) (D98G11) XP® Rabbit mAb.

对照(左)或λ 碱性磷酸酶处理(右)后,石蜡包埋的人宫颈癌组织,使用Phospho-NDRG1 (Thr346) (D98G11) XP® 兔单抗进行免疫组化分析。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-emebedded mouse colon using Phospho-NDRG1 (Thr346) (D98G11) XP® Rabbit mAb.

使用Phospho-NDRG1 (Thr346) (D98G11) XP® 兔单抗对石蜡包埋的小鼠结肠组织进行免疫组化分析。

Background

N-myc downstream-regulated gene 1 (NDRG1), also termed Cap43, Drg1, RTP/rit42, and Proxy-1, is a member of the NDRG family, which is composed of four members (NDRG1-4) that function in growth, differentiation, and cell survival (1-5). NDRG1 is ubiquitously expressed and highly responsive to a variety of stress signals including DNA damage (4), hypoxia (5), and elevated levels of nickel and calcium (2). Expression of NDRG1 is elevated in N-myc defective mice and is negatively regulated by N- and c-myc (1,6). During DNA damage, NDRG1 is induced in a p53-dependent fashion and is necessary for p53-mediated apoptosis (4,7). Research studies have shown that NDRG1 may also play a role in cancer progression by promoting differentiation, inhibiting growth, and modulating metastasis and angiogenesis (3,4,6,8,9). Nonsense mutation of the NDRG1 gene has been shown to cause hereditary motor and sensory neuropathy-Lom (HMSNL), which is supported by studies demonstrating the role of NDRG1 in maintaining myelin sheaths and axonal survival (10,11). NDRG1 is up-regulated during mast cell maturation and its deletion leads to attenuated allergic responses (12). Both NDRG1 and NDRG2 are substrates of SGK1, although the precise physiological role of SGK1-mediated phosphorylation is not known (13). NDRG1 is phosphorylated by SGK1 at Thr328, Ser330, Thr346, Thr356, and Thr366. Phosphorylation by SGK1 primes NDRG1 for phosphorylation by GSK-3.

N-myc下游调控gene1(NDRG1),也被称为Cap43,Drg1,RTP/rit42和Proxy-1,是NDRG家族的一员,该家族包含的4个成员(NDRG1-4)在生长,分化和细胞存活过程中发挥作用(1-5)。广泛表达的NDRG1并对多种压力信号发生应激反应,包括DNA损伤(4),缺氧(5),和镍及钙浓度升高(2)。NDRG1在N-myc缺陷小鼠中表达升高,能够被N-和c-myc负调控(1-6)。在DNA损伤过程中,NDRG1通过p53信号激活,并且是p53介导的凋亡过程中所必须的(4,7)。研究显示,NDRG1通过促进分化,抑制生长,调控血管生成和肿瘤转移等进程中发挥功能(3,4,6,8,9)。NDRG1基因发生无义突变后会引发遗传性运动和感觉神经病病变-Lom(HMSNL),该结论由证明NDRG1在维持髓鞘和轴突存活的研究支持(10,11)。NDRG1在肥大细胞成熟过程中表达上调,它的缺失会导致衰减的过敏反应(12)。NDRG1和NDRG2都是SGK1的底物,尽管SGK1介导的磷酸化的生理性作用尚未可知(13)。NDRG1可以被SGK1在Thr328, Ser330, Thr346, Thr356,以及Thr366磷酸化。NDRG1被SGK1磷酸化后可以被GSK-3磷酸化。

Phospho-NDRG1 (Thr346) (D98G11) XP® Rabbit mAb is directed at a site that was identified at Cell Signaling Technology (CST) using PhosphoScan®, CST's LC-MS/MS platform for modification site discovery. Phosphorylation at Thr346 was discovered using an Akt substrate antibody and was shown to be induced by insulin treatment in multiple cell lines. Please visit PhosphoSitePlus®, CST's modification site knowledgebase, at www.phosphosite.org for more information.

Cell Signaling Technology (CST)使用PhosphoScan®, CST's LC-MS/MS 平台以发现蛋白修饰点,设计了Phospho-NDRG1 (Thr346) (D98G11) XP® Rabbit mAb。Thr346的磷酸化是通过一个Akt底物抗体发现的,并在多种细胞系中发现可以被胰岛素刺激诱导表达。请访问www.phosphosite.org中的CST's修饰位点网站,PhosphoSitePlus®,以获得更多的信息。

  1. Shimono, A. et al. (1999) Mech Dev 83, 39-52.
  2. Zhou, D. et al. (1998) Cancer Res 58, 2182-9.
  3. van Belzen, N. et al. (1997) Lab Invest 77, 85-92.
  4. Kurdistani, S.K. et al. (1998) Cancer Res 58, 4439-44.
  5. Park, H. et al. (2000) Biochem Biophys Res Commun 276, 321-8.
  6. Li, J. and Kretzner, L. (2003) Mol Cell Biochem 250, 91-105.
  7. Stein, S. et al. (2004) J Biol Chem 279, 48930-40.
  8. Maruyama, Y. et al. (2006) Cancer Res 66, 6233-42.
  9. Nishio, S. et al. (2008) Cancer Lett 264, 36-43.
  10. Kalaydjieva, L. et al. (2000) Am J Hum Genet 67, 47-58.
  11. Okuda, T. et al. (2004) Mol Cell Biol 24, 3949-56.
  12. Taketomi, Y. et al. (2007) J Immunol 178, 7042-53.
  13. Murray, J.T. et al. (2004) Biochem J 384, 477-88.

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