WebFor simultaneous fixation and permeabilization of cells prior to intracellular staining Shop BD Phosflow™ Fix Buffer I at Fishersci.com WebFollowing fixation with 2% paraformaldehyde (PFA) and permeabilization with BD phosphoflow Perm buffer III, the mean fluo- rescence intensity of phospho-SMAD was quantified by flow cytometry, using anti-pSMAD2 (S465/467) SMAD3 (S423/425) rabbit mAb (Cell Signaling; D27F4), followed by anti-rabbit-PacificBlue secondary antibody.

BD™ Phosflow Protocols for Human PBMCs

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. WebDec 13, 2013 · After centrifugation at 300xg for 5 min, the supernatants were poured off and 1 ml -20°C cold BD Phosflow Perm Buffer III was added and the samples were incubated for 30 min in the dark on ice. The samples were then washed twice with 1 ml BD Phosphoflow stain buffer (centrifuged at 300x g for 5 min). ion television hawaii five-o

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WebPhosflow Perm Buffer II and Perm Buffer III, BD $215.00 / Each Catalog No. BDB554657 Bovine serum albumin Stain Buffer, BD $107.00 / Each Catalog No. BDB554722 BD Cytofix/Cytoperm™ Fixation and Permeabilizat $150.00 / Each Catalog No. BDB557885 Perm / Wash™ Buffer I (10x), BD $165.00 / Each Catalog No. BDB554723 BD … WebIntracellular Staining With True-Phos™ Perm Buffer in Cell Suspensions Protocol Protocol Steps Buffer Preparation: Warm Fixation Buffer (BioLegend Cat 420801 ). For each 1 x 10 6 cells aliquot 0.5mL of buffer and warm to 37°C. Chill True-Phos™ Perm Buffer to -20°C. For each 1 x 10 6 cells aliquot 1.0ml of True-Phos™ Perm Buffer and chill to -20°C. 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). on the green sports grill mesa