BD PharmingenTM PE Mouse anti-XBP-1S

Description

Multicolor flow cytometric analyses of XBP-1S expressed in human and mouse cells. Panel 1. Analysis of XBP-1S expressed in activated human peripheral blood mononuclear cells PBMC . Human PBMC were stimulated with 10 mu g/ml ODN 2006 Type B CpG Oligonucleotide Invivogen, Cat. No. tlrl-2006-1 at 37 C for 7 days. The cells were harvested and incubated with BD Horizon Fixable Viability Stain 450 Cat. No. 562247 , fixed in 1X Fix/Perm Buffer BD Pharmingen Transcription Buffer Set, Cat: 562574 at 2-8 C 45 min and permeabilized in 1X Perm/Wash Buffer Transcription Buffer Set at 2-8 C 45 min . Cells were stained with PE Mouse anti-XBP-1S Cat. No. 562642 and PerCP-Cy5.5 Mouse Anti-Human CD20 Cat. No. 558021 antibodies. Two-color flow cytometric contour plots showing the correlated expression of CD20 versus XBP-1S or Ig isotype control staining were derived from live-gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry was performed using a BD FACSCanto II Flow Cytometer System. Panel 2. Analysis of XBP-1S expressed in activated mouse splenocytes. BALB/C splenocytes were stimulated with 1 mu g/ml lipopolysaccharide LPS; Sigma Cat. No. L3137 at 37 C 3 days . Cells were incubated with Fixable Viability Stain 450, fixed in 1X Fix/Perm Buffer, and permeabilized in 1X Perm/Wash Buffer as described for Panel 1. The cells were stained with PE Mouse anti-XBP-1S and APC Rat Anti-Mouse CD45R/B220 BD Cat. No. 553092 . The cells were then analyzed as described above. Contour plots showing the correlated expression of B220 versus XBP-1S or Ig isotype control staining were derived from live-gated events with the forward and side light-scatter characteristics of intact lymphocytes. Multicolor flow cytometric analyses of XBP-1S expressed in human and mouse cells. Panel 1. Analysis of XBP-1S expressed in activated human peripheral blood mononuclear cells PBMC . Human PBMC were stimulated with 10 mu g/ml ODN 2006 Type B CpG Oligonucleotide Invivogen, Cat. No. tlrl-2006-1 at 37 C for 7 days. The cells were harvested and incubated with BD Horizon Fixable Viability Stain 450 Cat. No. 562247 , fixed in 1X Fix/Perm Buffer BD Pharmingen Transcription Buffer Set, Cat: 562574 at 2-8 C 45 min and permeabilized in 1X Perm/Wash Buffer Transcription Buffer Set at 2-8 C 45 min . Cells were stained with PE Mouse anti-XBP-1S Cat. No. 562642 and PerCP-Cy5.5 Mouse Anti-Human CD20 Cat. No. 558021 antibodies. Two-color flow cytometric contour plots showing the correlated expression of CD20 versus XBP-1S or Ig isotype control staining were derived from live-gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry was performed using a BD FACSCanto II Flow Cytometer System. Panel 2. Analysis of XBP-1S expressed in activated mouse splenocytes. BALB/C splenocytes were stimulated with 1 mu g/ml lipopolysaccharide LPS; Sigma Cat. No. L3137 at 37 C 3 days . Cells were incubated with Fixable Viability Stain 450, fixed in 1X Fix/Perm Buffer, and permeabilized in 1X Perm/Wash Buffer as described for Panel 1. The cells were stained with PE Mouse anti-XBP-1S and APC Rat Anti-Mouse CD45R/B220 BD Cat. No. 553092 . The cells were then analyzed as described above. Contour plots showing the correlated expression of B220 versus XBP-1S or Ig isotype control staining were derived from live-gated events with the forward and side light-scatter characteristics of intact lymphocytes. Show More Multicolor flow cytometric analyses of XBP-1S expressed in human and mouse cells. Panel 1. Analysis of XBP-1S expressed in activated human peripheral blood mononuclear cells PBMC . Human PBMC were stimulated with 10 mu g/ml ODN 2006 Type B CpG Oligonucleotide Invivogen, Cat. No. tlrl-2006-1 at 37 C for 7 days. The cells were harvested and incubated with BD Horizon Fixable Viability Stain 450 Cat. No. 562247 , fixed in 1X Fix/Perm Buffer BD Pharmingen Transcription Buffer Set, Cat: 562574 at 2-8 C 45 min and permeabilized in 1X Perm/Wash Buffer Transcription Buffer Set at 2-8 C 45 min . Cells were stained with PE Mouse anti-XBP-1S Cat. No. 562642 and PerCP-Cy5.5 Mouse Anti-Human CD20 Cat. No. 558021 antibodies. Two-color flow cytometric contour plots showing the correlated expression of CD20 versus XBP-1S or Ig isotype control staining were derived from live-gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry was performed using a BD FACSCanto II Flow Cytometer System. Panel 2. Analysis of XBP-1S expressed in activated mouse splenocytes. BALB/C splenocytes were stimulated with 1 mu g/ml lipopolysaccharide LPS; Sigma Cat. No. L3137 at 37 C 3 days . Cells were incubated with Fixable Viability Stain 450, fixed in 1X Fix/Perm Buffer, and permeabilized in 1X Perm/Wash Buffer as described for Panel 1. The cells were stained with PE Mouse anti-XBP-1S and APC Rat Anti-Mouse CD45R/B220 BD Cat. No. 553092 . The cells were then analyzed as described above. Contour plots showing the correlated expression of B220 versus XBP-1S or Ig isotype control staining were derived from live-gated events with the forward and side light-scatter characteristics of intact lymphocytes.