Flow cytometry

Flow cytometry general protocol

I. Required material Reagents - A primary antibody , against the target molecule - A Fc receptor blocker : « Anti-CD16/32, block Fc binding » that non-specifically binds primary antibodies - A secondary antibody in cases of indirect staining with a non labeled primary antibody - A non-relevant control isotype with a similar label than primary antibody Buffers - A staining buffer - An intracellular fixation buffer - Trypan blue...

Protocol for flow cytometry on whole blood

I. Required material Reagents - A primary antibody , against the target molecule - A secondary antibody in cases of indirect staining with a non labeled primary antibody - A non-relevant control isotype with a similar label than primary antibody Buffers - A staining buffer - A Red Blood Cells Lysis (RBC) buffer - An intracellular fixation buffer - Trypan blue for counting cells - Gaines sheath fluid Material - Pipets and Pipet-aid -...

Protocol for intracellular flow cytometry

I. Required material Reagents - A primary antibody against the target molecule - A secondary antibody in cases of indirect staining with a non labeled primary antibody - A non-relevant control isotype with a similar label than primary antibody Buffers - A staining buffer - A Permeabilization Buffer - An intracellular fixation buffer Material - Pipets and Pipet-aid - Centrifugator - Freezer - Flow cytometer II. Experimentation...