Nestin结构特征

Nestin为第VI类中间丝蛋白，其分子量为 220KD~250KD，位于1号染色体23.1q。Nestin基因含有三个内含子和四个外显子，三个内含子中有两个是与神经丝共享的，充分表明Nestin与神经丝来自于一个祖先的可能性。Nestin合成的蛋白分为三个区域，N端（1-7aa）、α螺旋区（8-313）和N端（314-1621aa），α螺旋区被分为四个部分（1A、1B、2A、2B）。

Nestin外显子与内含子分布结构图

Nestin结构分布图

Nestin主要功能

（1）参与细胞骨架形成，维持细胞形态；

（2）在有丝分裂过程中促进vimentin的解聚；

（3）维持脑与眼组织的正常发育；

（4）在一些肿瘤中，高表达nestin与肿瘤的浸润性生长、转移与不良预后成正相关。

Nestin临床应用

动物体早期发育过程中，中枢神经系统发育是一个重要事件。Nestin是一种中间丝蛋白，它在哺乳动物神经前体细胞中高表达，已被广泛用作神经前体细胞的标志分子。因此，nestin的表达情况对分析神经系统的进化具有重要作用，同时也可以作为神经系统病变和损伤的快速敏感诊断指标之一。在成体组织中，nestin只在部分具有分化潜能的细胞中表达，如在皮肤、心血管再生过程中表达增高。此外，nestin在多种肿瘤组织中表达增加，蛋白表达量与肿瘤的恶性程度成正相关，因此nestin在肿瘤组织中的表达水平对于评价肿瘤的生物学行为和患者的预后状况具有重要意义。

我们生产的nestin单克隆抗体（CSB-MA0157131A0m，100µg/50µg）可以应用于ELISA、WB、IHC、WB、IF、FC检测。WB可识别U251和SH-Sy5y天然样本；IHC检测显示在扁桃体、肾组织和黑色素瘤中为阳性；在Hela、PC-3、U251的IF检测中具有阳性信号；在hela和U251细胞的流式细胞检测中呈现强阳性。

WB:

Positive WB detected in：U251 whole cell lysate，SH-Sy5y whole cell lysate

All lanes: NES antibody at 0.6ug/ml

Predicted band size: 260 KDa

Observed band size: 260 KDa

IHC:

Immunohistochemistry of paraffin-embedded human tonsil tissue using CSB-MA0157131A0m at dilution of 1:100.

Immunohistochemistry of paraffin-embedded human kidney tissue using CSB-MA0157131A0m at dilution of 1:100.

Immunohistochemistry of paraffin-embedded human melanoma using CSB-MA0157131A0m at dilution of 1:100.

IF:

Immunofluorescent analysis of Hela cells using CSB-MA0157131A0m at a dilution of 1:100 and Alexa Fluor 488-congugated AffiniPure Goat Anti-Mouse IgG(H+L)

Immunofluorescent analysis of PC-3 cells using CSB-MA0157131A0m at a dilution of 1:100 and Alexa Fluor 488-congugated AffiniPure Goat Anti-Mouse IgG(H+L)

Immunofluorescent analysis of U251 cells using CSB-MA0157131A0m at a dilution of 1:100 and Alexa Fluor 488-congugated AffiniPure Goat Anti-Mouse IgG(H+L)

FC:

Overlay histogram showing U251 cells stained with CSB-MA0157131A0m (red line). The cells were fixed with 70% ethylalcohol (18h) and then permeabilized with 0.3% Triton X-100 for 2 min. The cells were then incubated in 1x PBS /10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (10µg/1x106cells) for 1 h at 4℃. The secondary antibody used was FITC goat anti-mouse IgG (H+L) at 1/200 dilution for 1 h at 4℃. Isotype control antibody (green line) was mouse IgG1 (10µg/1x106cells) used under the same conditions. Acquisition of >10,000 events was performed.

Overlay histogram showing Hela cells stained with CSB-MA0157131A0m (red line). The cells were fixed with 70% ethylalcohol (18h) and then permeabilized with 0.3% Triton X-100 for 2 min. The cells were then incubated in 1x PBS /10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (10µg/1x106cells) for 1 h at 4℃. The secondary antibody used was FITC goat anti-mouse IgG (H+L) at 1/200 dilution for 1 h at 4℃. Isotype control antibody (green line) was mouse IgG1 (10µg/1x106cells) used under the same conditions. Acquisition of >10,000 events was performed.