BD PhosflowTM Perm Buffer IV 10×

Description

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. 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. Show More 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.