Perm Buffer IV 10×(RUO)
Immunofluorescent staining of human and mouse lymphocytes after permeabilization with Perm Buffer IV. Human whole blood (top row) and mouse splenocytes (bottom row) were treated with 100 ng/ml IL-2 (Cat. No. 554603, top right figure) or 100 ng/ml IL-6 (Cat. No. 550071, bottom right figure) or left untreated (all 4 figures) for 15 minutes. Erythrocytes were lysed and leukocytes were fixed and permeabilized with Perm Buffer IV at either 1× or 0.5× according to the Recommended Assay Procedure. Then the human cells were stained with PE Mouse Anti-Human CD19 (Cat. No. 555413, top panel of top left figure), PE Mouse Anti-Human CD4 (Cat. No. 347327, bottom panel of top left figure), or PE Mouse Anti-Stat5 (pY694) (Cat. No. 612567, top right figure). Similarly, the mouse cells were stained with APC Hamster Anti-Mouse CD3e (Cat. No. 553066) and Alexa Fluor® 488 Rat Anti-Mouse CD45R (Cat. No. 557669) in the presence of Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™, Cat. No. 553142, bottom left figure) or PE Mouse Anti-Stat3 (pY705) (Cat. No. 612569, bottom right figure). Additional controls for the cell-surface staining were live cells that had undergone erythrocyte lysis using BD Pharm Lyse™ lysing buffer (Cat. No. 555899, left column) but no fixation or permeabilization. The fold change is indicated for staining of Stat5 (pY694) or Stat3 (pY705) with each perm buffer concentration (upper right corners of panels in the right column). Lymphocytes were selected by scatter profile. Flow cytometry was performed on a BD FACSCanto™ II flow cytometer.
BD™ Phosflow Perm Buffer IV 10×
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