Protocols | Flow Cytometry Core Facility Common dyes available that are quick and easy to use Prepare stock solution of each dye to 100X, and use in the following staining protocol (Ex If stock solution is 1mg mL, dilute 1 10 in PBS 1X) It is often a good idea to add viability dyes prior to analysis or sorting of samples
Key Steps in Flow Cytometry Protocols - MilliporeSigma Many antibodies used in flow cytometry are directly conjugated to a fluorochrome; however, many unlabeled primary antibodies are routinely used in combination with labeled secondary antibodies
Indirect flow cytometry (FACS) protocol - Abcam Indirect labelling requires two incubation steps, firstly with a primary antibody then with a compatible secondary antibody The secondary (and not the primary) antibody has the fluorescent dye (FITC, PE, Cy5, etc ) conjugated
How to Titrate Antibodies for Flow Cytometry - Biology Insights A common starting range might involve eight to ten dilutions, such as 1:25, 1:50, 1:100, 1:200, extending to dilutions like 1:1600 or 1:3200 This wide range captures the entire binding curve, ensuring the optimal concentration is not missed
Antibody Titration - Flow Cytometry Guide | Bio-Rad Titration requires dilutions of antibody to be made and the same number of cells stained in the same volume The dilution that represents the best stain index is the dilution to use
Protocol: Cell Surface Antibody Staining for Flow Cytometry This is our basic protocol for extracellular staining of cell surface epitopes in suspension cells for flow cytometry For intracellular staining, see our Protocol: Intracellular Antibody Staining for Flow Cytometry