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通过校准流式细胞术定量测定膜结合细胞因子受体。

Quantification of membrane-bound cytokine receptors by calibrated flow cytometry.

机构信息

Institute of Biology, Department of Systems Biology, Otto-von-Guericke University, 39106 Magdeburg, Germany.

Institute of Biology, Department of Systems Biology, Otto-von-Guericke University, 39106 Magdeburg, Germany; Center for Dynamic Systems: Systems Engineering (CDS), Otto-von-Guericke University, 39106 Magdeburg, Germany; Magdeburg Center for Systems Biology (MACS), Otto-von-Guericke University, 39106 Magdeburg, Germany; Center for Health and Medical Prevention (CHaMP), Otto-von-Guericke University, 39106 Magdeburg, Germany.

出版信息

STAR Protoc. 2023 Sep 15;4(3):102511. doi: 10.1016/j.xpro.2023.102511. Epub 2023 Aug 14.

DOI:10.1016/j.xpro.2023.102511
PMID:37581983
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10457439/
Abstract

We present a protocol for quantifying the expression of the receptor gp130 using a calibrated flow cytometric approach. We describe pitfalls for receptor quantification such as titration of primary antibodies and standardizing cell culture. Receptors are stained with primary antibodies and fluorophore-coupled secondary antibodies. Beads covered with defined numbers of immunoglobulin G stained with fluorophore-coupled secondary antibodies serve as calibrators. In this way, the fluorescence intensity of cells is converted to the number of receptors on the cell surface. For complete details on the use and execution of this protocol, please refer to Reeh et al. (2019)..

摘要

我们提出了一种使用校准流式细胞术定量测定受体 gp130 表达的方案。我们描述了受体定量测定中的一些陷阱，例如一抗的滴定和细胞培养的标准化。受体用一抗和荧光素偶联的二抗染色。用荧光素偶联的二抗染色的带有定义数量免疫球蛋白 G 的珠子作为校准物。通过这种方式，将细胞的荧光强度转换为细胞表面受体的数量。有关此方案的使用和执行的完整详细信息，请参阅 Reeh 等人。(2019)。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b1a2/10457439/a830d2dc318b/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b1a2/10457439/c36e9c359133/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b1a2/10457439/232664233fd9/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b1a2/10457439/41edefcdd598/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b1a2/10457439/c9efcf9eb71a/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b1a2/10457439/a830d2dc318b/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b1a2/10457439/c36e9c359133/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b1a2/10457439/232664233fd9/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b1a2/10457439/41edefcdd598/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b1a2/10457439/c9efcf9eb71a/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b1a2/10457439/a830d2dc318b/gr4.jpg

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