High mobility group protein B1 (HMGB1) belongs to a family of highly conserved proteins that contain HMG box domains. Human HMGB1 is expressed as a 215 amino acid (aa) single chain polypeptide containing three domains: two N-terminal globular, 70 aa positively charged DNA binding domains (HMG boxes A and B), and a negatively charged 30 aa C-terminal region that contains only Asp and Glu. Human HMGB1 is 100% aa identical to canine HMGB1 and 99% aa identical to mouse, rat, bovine and porcine HMGB1.
HMGB1 is a widely expressed and highly abundant protein. It was originally discovered as a nuclear protein that could bend DNA. Such bending stabilizes nucleosome formation and regulates the expression of select genes upon recruitment by DNA binding proteins. It is now known that HMGB1 also plays a significant role in extracellular signaling associated with inflammation. HMGB1 is massively released into the extracellular environment during cell necrosis. It acts as an inflammatory mediator that promotes monocyte migration and cytokine secretion, and as a mediator of T cell-dendritic cell interaction. In addition, activated monocytes, macrophages, and dendritic cells also secrete HMGB1, forming a positive feedback loop that results in the release of additional cytokines and neutrophils. The cytokine activity of HMGB1 is restricted to the HMG B box, while the A box is associated with the helix-loop-helix domain of transcription factors. Although HMGB1 does not possess a classic signal sequence, it appears to be secreted as an acetylated form via secretory endolysosome exocytosis. Once secreted, HMGB1 transduces cellular signals through its high affinity receptor, RAGE and, possibly, TLR2 and TLR4.
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不过也有用 HRP标记的protein a或g的，前提是你跑的样里没有IgG之类的东西。
2 一抗是属于哪个类或亚类
二抗需与一抗的类别或亚类相匹配。这通常是针对单克隆抗体而言。多克隆抗体主要是IgG类免疫球蛋白，因此相应的二抗就是抗IgG抗体.
单克隆抗体的类别及亚类通常会在产品列表中列出，如果你的一抗是小鼠IgM，那么相应的二抗就应当是抗小鼠IgM，或是抗小鼠IgG抗体。
如果单克隆一抗是小鼠IgG的某一亚类（IgG1，IgG2a，IgG2b，IgG3），那么几乎所有的抗小鼠IgG都可以与之结合，或者你也可以选择专门针对这一亚类的二抗，例如，如果你的一抗是小鼠IgG1，那么你可以选择抗IgG1的二抗，此种抗体在双标记实验中尤其适合。
如果你不知道一抗是哪一类别或亚类，那么抗小鼠IgG是一个不错的选择，因为此种抗体可以识别大多数类型的IgG免疫球蛋白。
