Cell Signaling Technology

Product Pathways - Cell Cycle / Checkpoint

Phospho-Cyclin D1 (Thr286) (D29B3) XP® Rabbit mAb #3300

CCND1   cyclind   cyclind1  

No. Size Price
3300S 100 µl ( 10 western blots ) ¥4,050.00 现货查询 购买询价
3300T 20 µl ( 2 western blots ) ¥1,600.00 现货查询 购买询价
3300 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Monkey, Endogenous 36 Rabbit IgG
IP 1:50
F 1:1600
IF-IC 1:2000

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, F=Flow Cytometry, IF-IC=Immunofluorescence (Immunocytochemistry),


Species predicted to react based on 100% sequence homology: Monkey,

Specificity / Sensitivity

Phospho-Cyclin D1 (Thr286) (D29B3) XP® Rabbit mAb detects endogenous levels of cyclin D1 only when phosphorylated at Thr286. The antibody does not cross-react with other cyclin D family members. Phospho-Cyclin D1 (Thr286) (D29B3) XP® Rabbit mAb能够检测内源性苏氨酸(286位点)磷酸化的cyclin D1蛋白,且不与cyclin D家族其他成员发生交叉反应。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr286 of cyclin D1. 该单克隆抗体是由合成的针对cyclin D1蛋白的苏氨酸(286位点)磷酸化肽段免疫动物生产的。

Western Blotting

Western Blotting

Western blot analysis of extracts from HT-1080 cells, untreated or treated with MG132 (10 μM, 4 hours), using Phospho-Cyclin D1 (Thr286) (D29B3) XP® Rabbit mAb. western blot方法分析未处理或MG132处理的 (10 μM, 4 hours)HT-1080细胞提取物,使用的抗体为Phospho-Cyclin D1 (Thr286) (D29B3) XP® Rabbit mAb。

Western Blotting

Western Blotting

Western blot analysis of extracts from HT-1080 cells, treated with MG132 and with or without λ phosphatase, using Phospho-Cyclin D1 (Thr286) (D29B3) XP® Rabbit mAb (upper) or Cyclin D1 (DC56) Mouse mAb #2926 (lower). western blot方法检测加入或未加入λ磷酸酶的情况下,经MG132处理的HT-1080细胞提取物,使用的抗体为Phospho-Cyclin D1 (Thr286) (D29B3) XP® Rabbit mAb(上图)或Cyclin D1 (DC56) Mouse mAb #2926 (下图)。

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of HT-1080 cells, untreated (blue) or MG132-treated (green), using Phospho-Cyclin D1 (Thr286) (D29B3) XP® Rabbit mAb. 流式细胞术分析未处理(蓝色)或MG-132处理的(绿色)HT-1080细胞,使用的抗体为Phospho-Cyclin D1 (Thr286) (D29B3) XP® Rabbit mAb。



Confocal immmunofluorescent analysis of HT-1080 (top) and Saos-2 cells (bottom), untreated (left) or treated with MG132 alone (center) or with MG132 followed by λ-phosphatase (right), using Phospho-Cyclin D1 (Thr286) (D29B3) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). 激光共聚焦荧光法检测HT-1080 (上) and Saos-2 cells (下), 左图为对照细胞,中间图为MG132单独处理,右图为MG132处理后加入λ磷酸酶,检测抗体为Phospho-Cyclin D1 (Thr286) (D29B3) XP® Rabbit mAb,呈绿色。肌动蛋白丝以DY-554 phalloidin标记,为红色。


Activity of the cyclin-dependent kinases CDK4 and CDK6 is regulated by T-loop phosphorylation, by the abundance of their cyclin partners (the D-type cyclins), and by association with CDK inhibitors of the Cip/Kip or INK family of proteins (1). The inactive ternary complex of cyclin D/CDK4 and p27 Kip1 requires extracellular mitogenic stimuli for the release and degradation of p27 concomitant with a rise in cyclin D levels to effect progression through the restriction point and pRb-dependent entry into S-phase (2). The active complex of cyclin D/CDK4 targets the retinoblastoma protein for phosphorylation, allowing the release of E2F transcription factors that activate G1/S-phase gene expression (3). Levels of cyclin D protein drop upon withdrawal of growth factors through downregulation of its protein expression and through phosphorylation-dependent degradation (4). 细胞周期蛋白依赖性蛋白激酶CDK4和CDK6的活性受三个因素的调控:T-环路磷酸化,周期蛋白伴侣(D-型周期蛋白)的丰度,与CDK抑制剂Cip/Kip 或 INK蛋白家族的关系(1)。失活的三元复合物cyclin D/CDK4 和p27 Kip1需要胞外有丝分裂刺激因子参与p27伴侣的释放和降解,伴随周期蛋白D的水平升高,通过限制点和pRb依赖性的S期进入影响细胞周期的进程(2)。活跃的cyclin D/CDK4复合体靶向视网膜母细胞瘤蛋白以利于磷酸化,促进可以激活G1/S期基因表达的转录因子E2F的释放(3)。生长因子退出后,通过下调蛋白表达和磷酸化依赖性蛋白降解,周期蛋白D的水平下降(4)。

Aberrant expression of cyclin D1 is associated with many forms of cancer, including B cell lymphomas. Gene translocation or amplification of the cyclin D1 gene can directly contribute to oncogenesis (2). Cyclin D1 also plays a critical role in mammary tissue maturation (5). Phosphorylation of cyclin D1 at Thr286 by glycogen synthase kinase 3β (4) or through the Ras/Raf/MEK/MAPK pathway (6) enhances its ubiquitination and proteasomal degradation. Cyclin D1的异常表达与许多形式的肿瘤相关,包括B细胞淋巴瘤。cyclin D1基因的转位或扩增可以直接促进肿瘤发生。Cyclin D1在乳腺组织成熟的过程中也起到了决定性的作用。通过GSK3或者Ras/Raf/MEK/MAPK途径使Cyclin D1 苏氨酸286位点磷酸化可以加强其泛素化和蛋白酶体降解。

  1. Hirai, H. et al. (1995) Mol Cell Biol 15, 2672-81.
  2. Sherr, C.J. (1996) Science 274, 1672-7.
  3. Lukas, J. et al. (1996) Mol Cell Biol 16, 6917-25.
  4. Diehl, J.A. et al. (1997) Genes Dev 11, 957-72.
  5. Sicinski, P. et al. (1995) Cell 82, 621-630.
  6. Shao, J. et al. (2000) J Biol Chem 275, 22916-24.

Application References

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