BD PhosflowTM Alexa Fluor 647 Mouse anti-WIP (pS488)

Description

Analysis of WIP pS488 in human T leukemia cells. LEFT: The shaded histogram displays Jurkat cells ATCC TIB152 that were stimulated by cross-linking of CD3 and CD28 with NA/LE Mouse anti-Human CD3 mAb Cat. No. 555329 and NA/LE Mouse anti-Human CD28 mAb Cat. No. 555725 on ice for 15 minutes followed by Purified Goat anti-Mouse Ig Cat. No. 553998 on ice for 15 minutes, and then allowed to undergo phosphorylation at 37C for 3 minutes. The open histogram shows unstimulated Jurkat cells. The cells were fixed using BD Cytofix buffer Cat. No. 554655 for 10 minutes at 37 C, permeabilized usingBD Phosflow Perm Buffer III Cat. No. 558050 on ice for at least 30 minutes, blocked with normal mouse immunoglobulin, and then stained with Alexa Fluor 647 Mouse anti-WIP pS488 . Flow cytometry was performed on a BD FACSArray bioanalyzer system. RIGHT: The specificity of mAb K32-824 was confirmed by western blot using unconjugated antibody on lysates from control left panel and CD3/CD28-cross-linked right panel Jurkat cells. WIP pS488 is upregulated in the treated cells; its observed molecular weight is 60 kDa, although the calculated molecular weight is 51 kDa. Analysis of WIP pS488 in human T leukemia cells. LEFT: The shaded histogram displays Jurkat cells ATCC TIB152 that were stimulated by cross-linking of CD3 and CD28 with NA/LE Mouse anti-Human CD3 mAb Cat. No. 555329 and NA/LE Mouse anti-Human CD28 mAb Cat. No. 555725 on ice for 15 minutes followed by Purified Goat anti-Mouse Ig Cat. No. 553998 on ice for 15 minutes, and then allowed to undergo phosphorylation at 37C for 3 minutes. The open histogram shows unstimulated Jurkat cells. The cells were fixed using BD Cytofix buffer Cat. No. 554655 for 10 minutes at 37 C, permeabilized usingBD Phosflow Perm Buffer III Cat. No. 558050 on ice for at least 30 minutes, blocked with normal mouse immunoglobulin, and then stained with Alexa Fluor 647 Mouse anti-WIP pS488 . Flow cytometry was performed on a BD FACSArray bioanalyzer system. RIGHT: The specificity of mAb K32-824 was confirmed by western blot using unconjugated antibody on lysates from control left panel and CD3/CD28-cross-linked right panel Jurkat cells. WIP pS488 is upregulated in the treated cells; its observed molecular weight is 60 kDa, although the calculated molecular weight is 51 kDa. Analysis of WIP pS488 in human T leukemia cells. LEFT: The shaded histogram displays Jurkat cells ATCC TIB152 that were stimulated by cross-linking of CD3 and CD28 with NA/LE Mouse anti-Human CD3 mAb Cat. No. 555329 and NA/LE Mouse anti-Human CD28 mAb Cat. No. 555725 on ice for 15 minutes followed by Purified Goat anti-Mouse Ig Cat. No. 553998 on ice for 15 minutes, and then allowed to undergo phosphorylation at 37C for 3 minutes. The open histogram shows unstimulated Jurkat cells. The cells were fixed using BD Cytofix buffer Cat. No. 554655 for 10 minutes at 37 C, permeabilized usingBD Phosflow Perm Buffer III Cat. No. 558050 on ice for at least 30 minutes, blocked with normal mouse immunoglobulin, and then stained with Alexa Fluor 647 Mouse anti-WIP pS488 . Flow cytometry was performed on a BD FACSArray bioanalyzer system. RIGHT: The specificity of mAb K32-824 was confirmed by western blot using unconjugated antibody on lysates from control left panel and CD3/CD28-cross-linked right panel Jurkat cells. WIP pS488 is upregulated in the treated cells; its observed molecular weight is 60 kDa, although the calculated molecular weight is 51 kDa. Show More Analysis of WIP pS488 in human T leukemia cells. LEFT: The shaded histogram displays Jurkat cells ATCC TIB152 that were stimulated by cross-linking of CD3 and CD28 with NA/LE Mouse anti-Human CD3 mAb Cat. No. 555329 and NA/LE Mouse anti-Human CD28 mAb Cat. No. 555725 on ice for 15 minutes followed by Purified Goat anti-Mouse Ig Cat. No. 553998 on ice for 15 minutes, and then allowed to undergo phosphorylation at 37C for 3 minutes. The open histogram shows unstimulated Jurkat cells. The cells were fixed using BD Cytofix buffer Cat. No. 554655 for 10 minutes at 37 C, permeabilized usingBD Phosflow Perm Buffer III Cat. No. 558050 on ice for at least 30 minutes, blocked with normal mouse immunoglobulin, and then stained with Alexa Fluor 647 Mouse anti-WIP pS488 . Flow cytometry was performed on a BD FACSArray bioanalyzer system. RIGHT: The specificity of mAb K32-824 was confirmed by western blot using unconjugated antibody on lysates from control left panel and CD3/CD28-cross-linked right panel Jurkat cells. WIP pS488 is upregulated in the treated cells; its observed molecular weight is 60 kDa, although the calculated molecular weight is 51 kDa. Show More Analysis of WIP pS488 in human T leukemia cells. LEFT: The shaded histogram displays Jurkat cells ATCC TIB152 that were stimulated by cross-linking of CD3 and CD28 with NA/LE Mouse anti-Human CD3 mAb Cat. No. 555329 and NA/LE Mouse anti-Human CD28 mAb Cat. No. 555725 on ice for 15 minutes followed by Purified Goat anti-Mouse Ig Cat. No. 553998 on ice for 15 minutes, and then allowed to undergo phosphorylation at 37C for 3 minutes. The open histogram shows unstimulated Jurkat cells. The cells were fixed using BD Cytofix buffer Cat. No. 554655 for 10 minutes at 37 C, permeabilized usingBD Phosflow Perm Buffer III Cat. No. 558050 on ice for at least 30 minutes, blocked with normal mouse immunoglobulin, and then stained with Alexa Fluor 647 Mouse anti-WIP pS488 . Flow cytometry was performed on a BD FACSArray bioanalyzer system. RIGHT: The specificity of mAb K32-824 was confirmed by western blot using unconjugated antibody on lysates from control left panel and CD3/CD28-cross-linked right panel Jurkat cells. WIP pS488 is upregulated in the treated cells; its observed molecular weight is 60 kDa, although the calculated molecular weight is 51 kDa. Analysis of WIP pS488 in human T leukemia cells. LEFT: The shaded histogram displays Jurkat cells ATCC TIB152 that were stimulated by cross-linking of CD3 and CD28 with NA/LE Mouse anti-Human CD3 mAb Cat. No. 555329 and NA/LE Mouse anti-Human CD28 mAb Cat. No. 555725 on ice for 15 minutes followed by Purified Goat anti-Mouse Ig Cat. No. 553998 on ice for 15 minutes, and then allowed to undergo phosphorylation at 37C for 3 minutes. The open histogram shows unstimulated Jurkat cells. The cells were fixed using BD Cytofix buffer Cat. No. 554655 for 10 minutes at 37 C, permeabilized usingBD Phosflow Perm Buffer III Cat. No. 558050 on ice for at least 30 minutes, blocked with normal mouse immunoglobulin, and then stained with Alexa Fluor 647 Mouse anti-WIP pS488 . Flow cytometry was performed on a BD FACSArray bioanalyzer system. RIGHT: The specificity of mAb K32-824 was confirmed by western blot using unconjugated antibody on lysates from control left panel and CD3/CD28-cross-linked right panel Jurkat cells. WIP pS488 is upregulated in the treated cells; its observed molecular weight is 60 kDa, although the calculated molecular weight is 51 kDa.