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β2-Microglobulin from human urine

β2 - 来自人尿的微球蛋白
Company: Sigma-Aldrich
Catalog#: M4890
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Peptide Loading on MHC Class I Molecules of Tumor Cells
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2016-09-20
[Abstract]  MHC class I molecules present peptides to cytotoxic T cells allowing the immune system to scan for intracellular pathogens and mutated proteins. The generation of antigenic peptides is a multistep process that ends in the endoplasmic reticulum (ER). Only peptides with the right length and sequence will bind nascent MHC class I molecules in the ER. This protocol allows for detachment of the endogenous peptides bound to MHC class I molecules by preserving them for the binding of high affinity synthetic peptides. The complete dissociation of endogenous peptides by mild acid treatment as well as the binding of synthetic peptides to MHC class I molecules will be evaluated measuring HLA class I molecules express on the cell surface by flow cytometry. The mouse antibody W6/32 which recognizes ... [摘要]  MHC I类分子将肽呈递到细胞毒性T细胞，允许免疫系统扫描胞内病原体和突变蛋白。抗原肽的产生是以内质网（ER）结束的多步过程。只有具有正确长度和序列的肽将结合ER中的新生MHC I类分子。该方案允许通过保持结合MHC I类分子的内源肽与高亲和力合成肽的结合而脱离。通过温和酸处理以及合成肽与MHC I类分子的结合来评价内源肽的完全解离，通过流式细胞术测量在细胞表面上表达的HLA I类分子。识别β2m相关HLA-A，-B，-C，-E和-G重链的小鼠抗体W6/32适合于此提议。表达表面HLA I类分子的任何肿瘤细胞系适于测定。另一个重要方面是知道肿瘤细胞系的HLA I类分型以允许选择已知的高亲和力肽。

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