| 货号 | 3875T |
| 描述 | The Aurora Antibody Sampler Kit provides an economical means to investigate the G2/M phase of the cell cycle. The kit contains enough primary and secondary antibodies to perform four Western blots with each antibody.Aurora Antibody Sampler Kit为研究细胞周期G2/M期提供了一个经济的方式。该试剂盒内含足够4次western blot实验的一抗和二抗。 |
| 目标/特异性 | Each antibody in the Aurora Antibody Sampler Kit detects endogenous levels of its respective target protein and does not cross-react with other family members. |
| 供应商 | CST |
| 背景 | Aurora kinases belong to a highly conserved family of mitotic serine/threonine kinases with three members identified among mammals: Aurora A, B, and C (1,2). Studies on the temporal expression pattern and subcellular localization of Aurora kinases in mitotic cells suggest an association with mitotic structure. Their functional influences span from G2 phase to cytokinesis and may be involved in key cell cycle events such as centrosome duplication, chromosome bi-orientation and segregation, cleavage furrow positioning, and ingression (3). Aurora A is detected at the centrosomes, along mitotic spindle microtubules, and in the cytoplasm of mitotically proliferating cells. Aurora A protein levels are low during G1 and S phases and peak during the G2/M phase of the cell cycle. Phosphorylation of Aurora A at Thr288 in its catalytic domain increases kinase activity. Aurora A is involved in centrosome separation, maturation, and spindle assembly and stability. Expression of Aurora B protein also peaks during the G2/M phase of the cell cycle; Aurora B kinase activity peaks at the transition from metaphase to the end of mitosis. Aurora B associates with chromosomes during prophase prior to relocalizing to the spindle at anaphase. Aurora B regulates chromosome segregation through the control of microtubule-kinetochore attachment and cytokinesis. Expression of both Aurora A and Aurora B during the G2/M phase transition is tightly coordinated with histone H3 phosphorylation (4,5); overexpression of these kinases is seen in a variety of human cancers (2,4). Aurora C localizes to the centrosome from anaphase to cytokinesis and both mRNA and protein levels peak during G2/M phase. Although typical Aurora C expression is limited to the testis, overexpression of Aurora C is detected in various cancer cell lines (6).极光激酶属于有丝分裂过程中高度保守的丝氨酸/苏氨酸激酶家族，在哺乳动物中已确定了三个成员：极光激酶A，极光激酶B和极光激酶C(1,2)。时空表达模式和有丝分裂细胞中极光激酶亚细胞定位的研究表明其和有丝分裂结构的关系。极光激酶功能的影响跨越G2期和细胞分裂期并可能涉及到关键的细胞周期事件，如中心体复制、染色体双向定位和隔离、卵裂沟定位和内移(3)。极光激酶A可以在中心体(沿纺锤体微管方向)和有丝分裂增殖细胞的胞质中被检测到。极光激酶A蛋白水平在G1和S期较低而在细胞周期的G2 / M期达峰值。极光激酶A催化结构域苏氨酸(288位)的磷酸化能够增强激酶的活性。极光激酶A参与中心体分离、成熟和纺锤体组装及稳定。极光激酶B的蛋白表达同样在细胞周期的G2/M期达峰值; 极光激酶B的激酶活性在分裂中期到有丝分裂结束的过渡阶段最高。在重新定位到有丝分裂后期纺锤体之前，极光激酶B和分裂前期的染色体密切相关。极光激酶B通过控制微管-着丝粒附着和细胞质分裂调节染色体分离。G2/M期过渡时极光激酶A和极光激酶B的表达紧密配合组蛋白H3的磷酸化(4,5); 多种人类癌症中都发现了这些激酶的过度表达(2,4)。极光激酶C集中于有丝分裂后期到细胞质分裂期的中心体中，而且其mRNA和蛋白质水平在G2 / M期达峰值。虽然典型的极光激酶C的表达仅限于睾丸，但是在各种肿瘤细胞株中都可以检测到其过度表达(6)。 |
| 存放说明 | -20C |
| 参考文献 | Warner, S.L. et al. (2003) Mol Cancer Ther 2, 589-95. Katayama, H. et al. (2003) Cancer Metastasis Rev 22, 451-64. Andrews, P.D. et al. (2003) Curr Opin Cell Biol 15, 672-83. Pascreau, G. et al. (2003) Prog Cell Cycle Res 5, 369-74. Crosio, C. et al. (2002) Mol Cell Biol 22, 874-85. Kimura, M. et al. (1999) J Biol Chem 274, 7334-40. |
Western blot analysis of extracts from HeLa, L929 and C6 cells, treated with 4 mM hydroxyurea for 20 hours or treated with 100 nM paclitaxel or 100 ng/ml nocodazole for 20 hours, using Phospho-Aurora A (Thr288)/Aurora B (Thr232)/Aurora C (Thr198) (D11A13) XP® Rabbit mAb #2914 (upper) or Aurora B/AIM1 Antibody #3094 (lower).Western blot方法检测HeLa, L929和C6细胞提取物，分为4 mM羟基脲处理20小时组、100 nM紫杉醇处理组和100 ng/ml诺考达唑处理20小时组，使用的抗体为Phospho-Aurora A (Thr288)/Aurora B (Thr232)/Aurora C (Thr198) (D11A13) XP® Rabbit mAb #2914 (上图) 或Aurora B/AIM1 Antibody #3094 (下图)。 | |
Western blot analysis of extracts from HeLa and HT29 cells, hydroxyurea or nocodazole treated, using Phospho-Aurora A (Thr288) (C39D8) Rabbit mAb #3079.Western blot检测羟基脲或诺考达唑处理的Hela和HT29细胞提取物，使用的抗体为 Phospho-Aurora A (Thr288) (C39D8) Rabbit mAb #3079。 | |
Confocal immunofluorescent analysis of HT-1080 cells using Phospho-Aurora A (Thr288)/Aurora B (Thr232)/Aurora C (Thr198) (D13A11) XP® Rabbit mAb (green), β-Tubulin (9F3) Rabbit mAb (Alexa Fluor® 555 Conjugate) #2116 (red), and Phospho-Histone H3 (Ser10) (6G3) Mouse mAb #9706 (blue). | |
Flow cytometric analysis of untreated Jurkat cells using Phospho-Aurora A (Thr288)/Aurora B (Thr232)/Aurora C (Thr198) (D13A11) XP® Rabbit mAb compared to propidium iodide (DNA content). The boxed population indicates phospho-Aurora A (Thr288)/Aurora B (Thr232)/Aurora C (Thr198)-positive cells. | |
Confocal immunofluorescent analysis of mitotic HeLa cells during metaphase (left) and anaphase (right) using Phospho-Aurora A (Thr288) (C39D8) Rabbit mAb (red) and Phospho-Histone H3 (Ser10) (6G3) Mouse mAb #9706 (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). | |
Western blot analysis of extracts from hydroxyurea or nocodazole treated HeLa and HT29 cells using Phospho-Aurora A (Thr288) (C39D8) Rabbit mAb. | |
Western blot analysis of extracts from HeLa, L929 and C6 cells, treated with 4 mM hydroxyurea for 20 hours to induce G1/S phase or treated with 100 nM paclitaxel or 100 ng/ml nocodazole for 20 hours to induce G2/M phase, using Phospho-Aurora A (Thr288)/Aurora B (Thr232)/Aurora C (Thr198) (D11A13) XP® Rabbit mAb (upper) or Aurora B/AIM1 Antibody #3094 (lower). | |
Western blot analysis of extracts from Hela, NIH/3T3 and C6 cells, untreated or nocodazole-treated (50 ng/ml, 24 hrs), using Aurora B/AIM1 Antibody. | |
Flow cytometric analysis of Hela cells, using Aurora B/AIM1 Antibody (blue) compared to a nonspecifc negative control antibody (red). | |
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO* is added and emits light during enzyme catalyzed decomposition. | |
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO* is added and emits light during enzyme catalyzed decomposition. | |
Immunoprecipitation of Aurora A from HeLa cell extracts using Rabbit (Da1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Aurora A (D3E4Q) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Aurora A (D3E4Q) Rabbit mAb. | |
Flow cytometric analysis of Jurkat cells using Aurora A (D3E4Q) Rabbit mAb and Propidium Iodide (PI)/RNase Staining Solution #4087 to measure DNA content. Anti-rabbit IgG (H+L), F(ab)2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody. | |
Western blot analysis of extracts from HT-29 cells, untreated (-) or synchronized in mitosis by treatment with thymidine (2 mM, 17 hr; +) and Nocodazole #2190 (100 ng/ml, 24 hr; +), using Aurora A (D3E4Q) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). | |
Confocal immunofluorescent analysis of HeLa cells using Aurora A (D3E4Q) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). |
