Revision 7
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IHC-P, IF-F, IF-IC, FC-FP

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

46, 48

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q92597

Entrez-Gene Id:

10397

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunohistochemistry (Paraffin) 1:250 - 1:1000
Immunofluorescence (Frozen) 1:50 - 1:100
Immunofluorescence (Immunocytochemistry) 1:200 - 1:800
Flow Cytometry (Fixed/Permeabilized) 1:200

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier-free (BSA and azide free) version of this product see product #89166.

Specificity / Sensitivity

Phospho-NDRG1 (Thr346) (D98G11) XP® Rabbit mAb 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.

Species Reactivity:

Human, Mouse, Rat, Monkey

Source / Purification

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

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 upregulated 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.

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.

  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.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IHC-P: Immunohistochemistry (Paraffin) IF-F: Immunofluorescence (Frozen) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品, (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Revision 7
#5482

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

Western Blotting Image 1: Phospho-NDRG1 (Thr346) (D98G11) XP® Rabbit mAb Expand Image
使用 Phospho-NDRG1 (Thr346) (D98G11) XP® Rabbit mAb (上图) 或总 NDRG1 Antibody #5196 (下图),对未经处理或经 LY294002 #9901、胰岛素处理或经这两者处理的 C2C12 细胞提取物进行蛋白质印迹分析。
Western Blotting Image 2: Phospho-NDRG1 (Thr346) (D98G11) XP® Rabbit mAb Expand Image
使用 Phospho-NDRG1 (Thr346) (D98G11) XP® Rabbit mAb (上图) 或总 NDRG1 Antibody #5196 (下图),对经牛小肠激酶和 λ 磷酸酶处理或未经处理的、 经 hEGF #8916(100ng/ml,30分钟) 处理的 A-431 细胞提取物进行蛋白质印迹分析。
Western Blotting Image 3: Phospho-NDRG1 (Thr346) (D98G11) XP® Rabbit mAb Expand Image
使用 Phospho-NDRG1 (Thr346) (D98G11) XP® Rabbit mAb (上图) 或 β-Tubulin (9F3) Rabbit mAb #2128 (下图),对hEGF #8916 (100 ng/ml, 30分钟) 处理、转染空载、或 SignalSilence® NDRG1 siRNA I #6245或 SignalSilence® NDRG1 siRNA II #6257转染的 Hela 细胞提取物进行蛋白质印迹分析。
Immunohistochemistry Image 1: Phospho-NDRG1 (Thr346) (D98G11) XP® Rabbit mAb Expand Image
使用 NDRG1 Antibody #5196(顶) 或 Phospho-NDRG1 (Thr346) (D98G11) XP® Rabbit mAb (底) 对 SignalSlide® Phospho-Akt (Ser473) IHC Controls #8101 [石蜡包埋的 LNCaP 细胞沉淀物、对照 (左图) 或经 LY294002 处理的 (右图)] 进行免疫组织化学分析。
Immunohistochemistry Image 2: Phospho-NDRG1 (Thr346) (D98G11) XP® Rabbit mAb Expand Image
使用 Phospho-NDRG1 (Thr346) (D98G11) XP® Rabbit mAb,对石蜡包埋的小鼠结肠进行免疫组织化学分析。
Immunohistochemistry Image 3: Phospho-NDRG1 (Thr346) (D98G11) XP® Rabbit mAb Expand Image
使用 Phospho-NDRG1 (Thr346) (D98G11) XP® Rabbit mAb,对于对照 (左图) 或经 λ 磷酸酶处理 (右图) 的石蜡包埋人子宫颈癌进行免疫组织化学分析。
Immunohistochemistry Image 4: Phospho-NDRG1 (Thr346) (D98G11) XP® Rabbit mAb Expand Image
使用 Phospho-NDRG1 (Thr346) (D98G11) XP® Rabbit mAb 对石蜡包埋的人类肺癌进行免疫组织化学分析。
Immunofluorescence Image 1: Phospho-NDRG1 (Thr346) (D98G11) XP® Rabbit mAb Expand Image
对使用 Phospho-NDRG1 (Thr346) (D98G11) XP® Rabbit mAb #5482(绿色)标记的固定冷冻的小鼠结肠组织进行共聚焦免疫荧光分析。随后用 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 封闭游离的二抗结合位点,之后用 α-Smooth Muscle Actin (D4K9N) XP® Rabbit mAb (Alexa Fluor® 555 Conjugate) #60839(黄色)、DyLight 650 Phalloidin #12956(红色)和 DAPI #4083(蓝色)进行共标记。
Immunofluorescence Image 2: Phospho-NDRG1 (Thr346) (D98G11) XP® Rabbit mAb Expand Image
对使用 Phospho-NDRG1 (Thr346) (D98G11) XP® Rabbit mAb #5482(左图,绿色)标记并使用 DyLight 650 Phalloidin #12956(右图,红色)和 DAPI #4083(右图,蓝色)共标记的固定冷冻的小鼠肾脏组织进行共聚焦免疫荧光分析。
Immunofluorescence Image 3: Phospho-NDRG1 (Thr346) (D98G11) XP® Rabbit mAb Expand Image
对使用 Phospho-NDRG1 (Thr346) (D98G11) XP® Rabbit mAb #5482(绿色)标记的固定冷冻的小鼠纹状体组织进行共聚焦免疫荧光分析。使用 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 封闭游离的二抗结合位点,之后用 Myelin Basic Protein (D8X4Q) XP® Rabbit mAb (Alexa Fluor® 555 Conjugate) #84987(黄色)、GFAP (GA5) Mouse mAb (Alexa Fluor® 647 Conjugate) #3657(红色)和 DAPI #4083(蓝色)进行共标记。
Immunofluorescence Image 1: Phospho-NDRG1 (Thr346) (D98G11) XP® Rabbit mAb Expand Image
使用 Phospho-NDRG1 (Thr346) (D98G11) XP® Rabbit mAb (绿色),对经 LY294002 #9901处理 (左图) 或胰岛素处理 (右图) 的 C2C12 细胞进行共聚焦免疫荧光分析。使用 DY-554 phalloidin(红色)标记肌动蛋白纤丝。蓝色伪彩 = DRAQ5® #4084(DNA 荧光染料)。
Flow Cytometry Image 1: Phospho-NDRG1 (Thr346) (D98G11) XP® Rabbit mAb Expand Image

使用 Phospho-NDRG1 (Thr346) (D98G11) XP® Rabbit mAb(实线)或浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(虚线)对未经处理(绿色)或经 LY294002 #9901 (50 µM)、Wortmannin #9951 (1 µM) 和 U0126 #9903 (10 μM)(蓝色)处理 2 小时的 Jurkat 细胞进行流式细胞分析。Anti-rabbit IgG (H+L)、F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 作为二抗。