信帆生物：細(xì)胞因子類(lèi)ELISA試劑盒文獻(xiàn)上線

《P-selectin glycoprotein ligand 1 deficiency prevents development of acute pancreatitis by attenuating leukocyte infiltration》

摘要：

BACKGROUND Acute pancreatitis(AP) is rapid-onset pancreatic inflammation that causes local and systemic inflammatory response syndrome(SIRS) with high morbidity and mortality, but no approved therapies are currently available. P-selectin glycoprotein ligand 1(PSGL-1) is a transmembrane glycoprotein to initiate inflammatory responses. We hypothesized that PSGL-1 may be involved in the development of AP and would be a new target for the treatment of AP.AIM To investigate the role and mechanism of PSGL-1 in the development of AP.METHODS The PSGL-1 expression on leukocytes was detected in peripheral blood of AP patients and volunteers. Pancreatic injury, inflammatory cytokines expression, and inflammatory cell infiltration was measured in AP mouse models induced with PSGL-1 knockout(PSGL-1-/-) and wild-type(PSGL-1+/+) mice. Leukocyteendothelial cell adhesion was measured in a peripheral blood mononuclear cell(PBMC)-endothelial cell coculture system.RESULTS The expression of PSGL-1 on monocytes and neutrophils was significantly increased in AP patients. 

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二、操作步驟：

1. 從室溫平衡20min后的鋁箔袋中取出所需板條，剩余板條用自封袋密封放回4℃。

2. 設(shè)置標(biāo)準(zhǔn)品孔和樣本孔，標(biāo)準(zhǔn)品孔各加不同濃度的標(biāo)準(zhǔn)品50μL。

3. 樣本孔中加入待測(cè)樣本50μL；空白孔不加。

4. 除空白孔外，標(biāo)準(zhǔn)品孔和樣本孔中每孔加入辣根過(guò)氧化物酶（HRP）標(biāo)記的檢測(cè)抗體100μL，用封板膜封住反應(yīng)孔，37℃水浴鍋或恒溫箱溫育60min。

5. 棄去液體，吸水紙上拍干，每孔加滿(mǎn)洗滌液（350μL），靜置1min，甩去洗滌液，吸水紙上拍干，如此重復(fù)洗板5次（也可用洗板機(jī)洗板）。

6. 每孔加入底物A、B各50μL，37℃避光孵育15min。

7. 每孔加入終止液50μL，15min內(nèi)，在450nm波長(zhǎng)處測(cè)定各孔的OD值。 

三、使用安全措施：

請(qǐng)務(wù)必穿戴好實(shí)驗(yàn)服、防護(hù)面罩和實(shí)驗(yàn)手套等，做好實(shí)驗(yàn)防護(hù)。嚴(yán)禁直接接觸產(chǎn)品。剩余產(chǎn)品請(qǐng)勿隨意丟棄，請(qǐng)按照實(shí)驗(yàn)室相關(guān)生物和化學(xué)廢棄物的處理規(guī)定，進(jìn)行處理。