如何體外檢測(cè)高遷移率族蛋白1
高遷移率族蛋白1（High Mobility Group Protein 1），也被稱為二性霉素，由HMGB1基因編碼，由于在聚丙烯酰胺凝膠電泳中有很高的遷移率而被命名為高遷移率族蛋白1。
下面我們主要以人HMGB1為參考，對(duì)高遷移率族蛋白1進(jìn)行簡(jiǎn)要的介紹。
蛋白：
HMGB1跟組蛋白一樣，HMGB1是最重要的染色質(zhì)蛋白，在維持核小體穩(wěn)定及DNA重組、復(fù)制、修復(fù)及基因轉(zhuǎn)錄扮演重要角色。此外，它在感染、炎癥及免疫反應(yīng)中也發(fā)揮重要的作用。HMGB1也是炎癥損傷的研究熱點(diǎn)之一。已有學(xué)者提出將HMGB1作為DNA疫苗的佐劑和癌癥治療的靶標(biāo)。
根據(jù)Uniprot上人HMGB1 鏈接，http://www.uniprot.org/uniprot/P09429，我們制備了三種HMGB1片段，序列選取分別是Met1~Glu215(RPA399Hu01)，Pro9~Arg163（RPA399Hu02），Met1~Glu215（RPA399Hu61）。RPA399Hu01和RPA399Hu01是E.coli中表達(dá)的，而RPA399Hu61用的是293F進(jìn)行表達(dá)。這樣研究者可根據(jù)自己實(shí)驗(yàn)?zāi)康?，選用不同的片段以及表達(dá)系統(tǒng)。
抗體：
獲得制備的蛋白后，免疫小鼠和兔，獲得單克隆抗體和多克隆抗體后經(jīng)過(guò)親和層析純化得到高純度的抗體。單克隆抗體MAA399Hu22，該抗體效價(jià)為81w 。
試劑盒:
獲得了單克隆抗體和多克隆抗體后，我們研發(fā)了HMG1的Elisa試劑盒。試劑盒中包被的是小鼠單克隆抗體，檢測(cè)抗體是兔多克隆抗體。HMG1在正常和不同疾病模型中的濃度會(huì)有很大的變化，武漢云克隆公司也檢測(cè)了許多樣本，根據(jù)內(nèi)部實(shí)驗(yàn)結(jié)果，我們對(duì)46例表觀健康的人進(jìn)行了檢測(cè)，HMG1的濃度范圍為72~582ng/ml，僅供參考。
另外，僅僅是檢測(cè)試劑盒SEA399Hu，我們目前擁有的文獻(xiàn)就有24篇，研究者可以放心使用。
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