WebThe phosphoflow process to measure STAT-3 and STAT-5 included the lysis and fixation, washing, and subsequent permeabilization of the cells for 30 minutes at 4°C. The cells … WebThe phosphoflow process to measure STAT-3 and STAT-5 included the lysis and fixation, washing, and subsequent permeabilization of the cells for 30 minutes at 4°C. The cells were then stained for the surface markers CD3, CD4, CD8, and CD19 for one hour at room temperature, before acquisition on a BD Fortessa™.
Transcription Factor Phospho Buffer Set - BD Biosciences
WebMay 9, 2013 · Then, cells were fixed with Cytofix Buffer (BD Biosciences) and permeabilized in chilled Phosphoflow PermBuffer-III for 30 minutes before staining with anti-extracellular signal-regulated kinase (ERK)1/2 (pT302/pY204)-PE, anti-STAT5 (pY694)-PE, and anti-Akt (also known as protein kinase B; pS473). WebPhosphoflow is commonly performed on whole blood, but it can be performed on platelet-rich plasma (PRP), washed platelets, and murine samples with comparable ease.11In any case, blood should be drawn into an anticoagulant tube (e.g., BD Vacutainer), and sample prepara- tion should begin as soon as possible.7Assays are often performed on iphone xr refurb
Phosphoflow Cytometry and Barcoding in Blood Platelets: …
WebMay 15, 2024 · Phosphoflow cytometry (phosphoflow) is a single-cell-based technique to measure phosphorylated intracellular proteins, providing a more quantitative read-out than Western blotting. Recent advances in phosphoflow basically allow simultaneous analysis of protein phosphorylation in B cell (sub)populations, without prior cell sorting. However ... WebBD Phosflow™ Perm Buffer III is the recommended permeabilization buffer for the detection of phosphorylated proteins by flow cytometry. Three alternate permeabilization buffers are also available and may be useful for detection of a particular phosphoprotein or for phosphoproteins in combination with surface or other intracellular proteins. WebProtocol III Harsh alcohol method Protocol IV Detergent method Fix buffer recommended for PBMCs or cell lines: BD Cytofix™ Buffer (554655) BD Cytofix™ Buffer (554655) BD … orange theory johnson city