Simultaneous blockade of Fc gamma receptors and indirect labeling of mouse lymphocytes by the selective detection of allotype-restricted epitopes on the kappa chain of rat monoclonal antibodies.
Simultaneous blockade of Fc gamma receptors and indirect labeling of mouse lymphocytes by the selective detection of allotype-restricted epitopes on the kappa chain of rat monoclonal antibodies.
复制标题
通过选择性检测大鼠单克隆抗体 kappa 链上的同种异型限制性表位，同时阻断 Fc γ 受体并间接标记小鼠淋巴细胞。
DOI：
10.1002/cyto.10057
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发表时间：
2002
期刊：
影响因子：
--
通讯作者：
Szakal,AndrasK
中科院分区：
文献类型：
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作者：
Balogh,Peter;Tew,JohnG;Szakal,AndrasK
背景将小鼠造血细胞与 IgG 类大鼠单克隆抗体 (mAb) 一起孵育有时会导致 Fc 区介导的 Fc 受体与免疫球蛋白的结合。这种不需要的结合可以通过将靶细胞与大鼠抗小鼠抗 CD16/32 (2.4G2) mAb 预孵育来防止。方法为了避免荧光染料偶联的二抗大鼠抗体与 Fc 受体阻断 2.4G2 发生交叉反应，需要直接荧光染料偶联的免疫球蛋白 用过的。然而，我们报告说，具有强烈大鼠 Igka 同种异型偏好的小鼠单克隆抗体 (MRC OX12) 可与未标记的大鼠单克隆抗体结合使用进行流式细胞术测量，并同时通过 2.4G2 单克隆抗体阻断 Fcγ 受体。 结果和讨论 OX12 对 2.4G2 单克隆抗体缺乏反应性，这是值得注意的，因为它可以有效地检测另一种单克隆抗体。 Sprague-Dawley 衍生的大鼠单克隆抗体。这种染色程序（在 2.4G2 IgG 存在的情况下，通过 OX12 mAb 检测适当菌株的未标记大鼠 mAb）是使用直接抗体缀合物的一种有吸引力的替代方案，同时满足有效 Fcγ 受体阻断的需要。细胞计数 47:107–110, 2002。© 2002 Wiley-Liss, Inc.
BackgroundIncubation of mouse hemopoietic cells with rat monoclonal antibodies (mAbs) of the IgG class sometimes results in Fc‐region mediated binding of immunoglobulins by Fc‐receptors. This unwanted binding can be prevented by preincubation of target cells with the rat anti‐mouse anti‐CD16/32 (2.4G2) mAb.MethodsTo avoid the cross‐reactivity of fluorochrome‐conjugated secondary anti‐rat antibodies with the Fc‐receptor blocking 2.4G2, direct fluorochrome‐conjugated immunoglobulins need to be used. However, we report that a mouse mAb (MRC OX12) with a strong rat Igkaallotype preference can be used for flow cytometric measurements in conjunction with unlabeled rat mAbs with the simultaneous blockade of Fcγ receptors by the 2.4G2 mAb.Results and DiscussionThis lack of reactivity of OX12 against the 2.4G2 mAb is remarkable, as it could efficiently detect another Sprague‐Dawley–derived rat mAb. This staining procedure (unlabeled rat mAb of the appropriate strain detected by OX12 mAb in the presence of 2.4G2 IgG) is an attractive alternative to using direct antibody conjugates, while satisfying the need for an effective Fcγ‐receptor blockade. Cytometry 47:107–110, 2002. © 2002 Wiley‐Liss, Inc.