| 货号 | 12007S |
| 描述 | This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 488 fluorescent dye and tested in-house for direct immunofluorescent analysis in human cells. This antibody is expected to exhibit the same species cross-reactivity as the unconjugated Glucocorticoid Receptor (D8H2) XP® Rabbit mAb #3660.该Cell Signaling Technology抗体与Alexa Fluor® 488荧光染料结合，并直接用流式细胞术内部测试检测人体细胞。预计该抗体与非结合的Glucocorticoid Receptor (D8H2) XP® Rabbit mAb #3660一样，表现出相同的物种交叉反应。 |
| 反应种属 | Human/Mouse/Rat/Monkey |
| 来源宿主 | Rabbit |
| 应用 | IF-IC/F |
| 使用方法 | F(1:100) IF-IC (1:50) |
| 供应商 | CST |
| 标记 | Alexa Fluor 488 |
| 背景 | Glucocorticoid hormones control cellular proliferation, inflammation, and metabolism through their association with the glucocorticoid receptor (GR)/NR3C1, a member of the nuclear hormone receptor superfamily of transcription factors (1). GR is composed of several conserved structural elements, including a carboxy-terminal ligand-binding domain (which also contains residues critical for receptor dimerization and hormone-dependent gene transactivation), a neighboring hinge region containing nuclear localization signals, a central zinc-finger-containing DNA-binding domain, and an amino-terminal variable region that participates in ligand-independent gene transcription. In the absence of hormone, a significant population of GR is localized to the cytoplasm in an inactive form via its association with regulatory chaperone proteins, such as HSP90, HSP70, and FKBP52. On hormone binding, GR is released from the chaperone complex and translocates to the nucleus as a dimer to associate with specific DNA sequences termed glucocorticoid response elements (GREs), thereby enhancing or repressing transcription of specific target genes (2). It was demonstrated that GR-mediated transcriptional activation is modulated by phosphorylation (3-5). Although GR can be basally phosphorylated in the absence of hormone, it becomes hyperphosphorylated upon binding receptor agonists. It has been suggested that hormone-dependent phosphorylation of GR may determine target promoter specificity, cofactor interaction, strength and duration of receptor signaling, receptor stability, and receptor subcellular localization (3). Indeed Ser211 of human GR is phosphorylated to a greater extent in the presence of hormone, and biochemical fractionation studies following hormone treatment indicate that Ser211-phosphorylated GR is found in the nucleus (3). Thus, Ser211 phosphorylation is a biomarker for activated GR in vivo. An added layer of complexity to GR signaling lies in the ability of multiple isoforms to be generated through both alternative splicing and the use of alternative translation intiation start sites, thus increasing the repertoire of functional signaling homo- and heterodimers (6,7). 糖皮质激素通过与转录因子的核激素受体超家族成员——糖皮质激素受体（GR）/ NR3C1联合，控制细胞增殖、炎症和代谢（1）。GR是由一些保守的结构元件，包括一个COOH-末端配体结合域（其中还包含受体二聚化和激素依赖型基因转录的关键残基），一个含有核定位信号的相邻的铰链区，一个含DNA结合结构域的中央锌指和一个参与配体无关的基因转录的NH 2 - 末端可变区。在没有激素的情况下，一个明显的GR类群以无活性方式通过其监管伴侣蛋白，如热休克蛋白90、热休克蛋白70和FKBP52，定位到细胞质中。对于激素结合，GR从分子伴侣复合物释放并作为二聚体转移到细胞核，与特定的DNA序列一起被称为糖皮质激素反应元件（GREs），增加或抑制特定的靶基因转录（2）。据证明，GR-介导的转录激活受磷酸化调节（3-5）。虽然GR可以在没有激素的情况下初级磷酸化。建议GR激素依赖型磷酸化可能决定靶启动子的特异性、辅助因子相互作用、受体信号的强度和持续时间、受体的稳定性和受体的亚细胞定位（3）。事实上，GR磷酸化更大程度上是在激素和生化分馏研究存在的情况下，可在细胞核中发现激素治疗表明丝氨酸（211位）-磷酸化GR（3）。因此，丝氨酸（211）磷酸化是体内激活GR的生物标志物。GR信号附加层的复杂性在于通过选择性剪接和使用选择性翻译启动起始位点产生多发亚型的能力，从而增加了功能信号同源和异源二聚体库（6,7）。 |
| 存放说明 | 4C |
Confocal immunofluorescent analysis of HeLa cells, grown in phenol red-free media containing 5% charcoal-stripped FBS for 2 d, and either untreated (left) or dexamethasone-treated (100 nM, 2 hr; right), using Glucocorticoid Receptor (D8H2) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate) (green). Actin filaments were labeled with DY-554 phalloidin (red).激光共聚焦免疫荧光法检测HeLa细胞，细胞培养在不含酚红，含5%活性碳/葡聚糖处理FBS中，之后细胞不处理（左图）或用地塞米松处理（100 nM, 2 hr; 右图），使用的抗体为Glucocorticoid Receptor (D8H2) XP® Rabbit mAb (Alexa Fluor® 488 结合) (绿色).肌动蛋白丝用DY-554鬼笔环肽标记（红色）。 | |
Human whole blood was fixed, lysed, and permeabilized as per the Cell Signaling Technology Flow Cytometry (Alternate) Protocol and stained with CD3-PE, CD19-APC and Glucocorticoid Receptor (D8H2) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate). CD3 (blue) and CD19 (green) population gates were applied to a histogram depicting the mean fluorescence intensity of glucocorticoid and compared to Rabbit (DA1E) mAb IgG XP® Isotype Control (Alexa Fluor® 488 Conjugate) #2975 (red). |
