Nestin結(jié)構(gòu)特征

Nestin為第VI類中間絲蛋白，其分子量為 220KD~250KD，位于1號(hào)染色體23.1q。Nestin基因含有三個(gè)內(nèi)含子和四個(gè)外顯子，三個(gè)內(nèi)含子中有兩個(gè)是與神經(jīng)絲共享的，充分表明Nestin與神經(jīng)絲來(lái)自于一個(gè)祖先的可能性。Nestin合成的蛋白分為三個(gè)區(qū)域，N端（1-7aa）、α螺旋區(qū)（8-313）和N端（314-1621aa），α螺旋區(qū)被分為四個(gè)部分（1A、1B、2A、2B）。

 

Nestin外顯子與內(nèi)含子分布結(jié)構(gòu)圖

 

Nestin結(jié)構(gòu)分布圖

Nestin主要功能

（1）參與細(xì)胞骨架形成，維持細(xì)胞形態(tài)；

（2）在有絲分裂過(guò)程中促進(jìn)vimentin的解聚；

（3）維持腦與眼組織的正常發(fā)育；

（4）在一些腫瘤中，高表達(dá)nestin與腫瘤的浸潤(rùn)性生長(zhǎng)、轉(zhuǎn)移與不良預(yù)后成正相關(guān)。

Nestin臨床應(yīng)用

動(dòng)物體早期發(fā)育過(guò)程中，中樞神經(jīng)系統(tǒng)發(fā)育是一個(gè)重要事件。Nestin是一種中間絲蛋白，它在哺乳動(dòng)物神經(jīng)前體細(xì)胞中高表達(dá)，已被廣泛用作神經(jīng)前體細(xì)胞的標(biāo)志分子。因此，nestin的表達(dá)情況對(duì)分析神經(jīng)系統(tǒng)的進(jìn)化具有重要作用，同時(shí)也可以作為神經(jīng)系統(tǒng)病變和損傷的快速敏感診斷指標(biāo)之一。在成體組織中，nestin只在部分具有分化潛能的細(xì)胞中表達(dá)，如在皮膚、心血管再生過(guò)程中表達(dá)增高。此外，nestin在多種腫瘤組織中表達(dá)增加，蛋白表達(dá)量與腫瘤的惡性程度成正相關(guān)，因此nestin在腫瘤組織中的表達(dá)水平對(duì)于評(píng)價(jià)腫瘤的生物學(xué)行為和患者的預(yù)后狀況具有重要意義。

 

我們生產(chǎn)的nestin單克隆抗體（CSB-MA0157131A0m，100µg/50µg）可以應(yīng)用于ELISA、WB、IHC、WB、IF、FC檢測(cè)。WB可識(shí)別U251和SH-Sy5y天然樣本；IHC檢測(cè)顯示在扁桃體、腎組織和黑色素瘤中為陽(yáng)性；在Hela、PC-3、U251的IF檢測(cè)中具有陽(yáng)性信號(hào)；在hela和U251細(xì)胞的流式細(xì)胞檢測(cè)中呈現(xiàn)強(qiáng)陽(yáng)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.