兔單克隆抗體 to Nestin

Rabbit

適用于: ICC/IF, Flow Cyt, WB, IHC-P

與反應(yīng): Human
預(yù)測可用于: Dog

Synthetic peptide within Human Nestin aa 1600 to the C-terminus (C terminal). The exact sequence is proprietary.
Database link: P48681

Western blot - Anti-Nestin antibody [SP103] (ab105389)

All lanes : Anti-Nestin antibody [SP103] (ab105389) at 1/100 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : NES knockout HAP1 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 177 kDa
Lanes 1 - 2: Merged signal (red and green). Green - ab105389 observed at 300 kDa. Red - loading control, ab18058, observed at 117 kDa.
ab105389 was shown to specifically react with NES in wild-type HAP1 cells as signal was lost in NES knockout cells. Wild-type and NES knockout samples were subjected to SDS-PAGE. Ab105389 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/100 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

ANTI-NESTIN抗體是一類可以特異性結(jié)合ANTI-NESTIN的多克隆抗體，主要用于檢測ANTI-NESTIN的Western Blot、IHC-P、IF、ELISA、Co-IP等多種免疫學(xué)實(shí)驗(yàn)。

雙抗體夾心法測定標(biāo)本中ANTI-NESTIN水平。用純化的ANTI-NESTIN抗體包被微孔板，制成固相抗體，往包被單抗的微孔中依次加入ANTI-NESTIN，再與HRP標(biāo)記的ANTI-NESTIN抗體結(jié)合，形成抗體-抗原-酶標(biāo)抗體復(fù)合物，經(jīng)過徹底洗滌后加底物TMB顯色。TMB在HRP酶的催化下轉(zhuǎn)化成藍(lán)色，并在酸的作用下轉(zhuǎn)化成最終的黃色。顏色的深淺和樣品中的ANTI-NESTIN呈正相關(guān)。用酶標(biāo)儀在450nm波長下測定吸光度（OD值），通過標(biāo)準(zhǔn)曲線計(jì)算樣品中ANTI-NESTIN濃度。

Nestin是一種屬于第Ⅵ類中間絲的細(xì)胞骨架蛋白,表達(dá)于多種組織的干細(xì)胞或前體細(xì)胞中,被廣泛用作干細(xì)胞的標(biāo)志性蛋白。除維持細(xì)胞的正常形態(tài),nestin還具有抗凋亡作用。抑制細(xì)胞中nestin的表達(dá),可以增強(qiáng)過氧化氫(H202)刺激誘導(dǎo)的神經(jīng)前體細(xì)胞和血管平滑肌細(xì)胞的凋亡,而將野生型的nestin(nest-1893)轉(zhuǎn)入到細(xì)胞內(nèi)可起到保護(hù)作用,顯著減少H202誘導(dǎo)的細(xì)胞凋亡。