CD154 MicroBead Kit, human

Name: CD154 MicroBead Kit, human
Availability: InStock

CD154 MicroBead Kit, human

The CD154 MicroBead Kit has been developed for positive selection of activated CD154

⁺

antigen-specific CD4

⁺

T cells from human PBMCs.

Product data and images for CD154 MicroBead Kit, human

Figures

Figure 1

CD154

⁺

T cells were isolated from human PBMCs of a CMV

⁺

donor using the CD154 MicroBead Kit, two MS Columns, and a MiniMACS™ Separator. PBMCs were stimulated for 16 hours with PepTivator CMV pp65 (# 130-093-438), a CD40 blocking-antibody was added during the stimulation to prevent down-regulation of CD154. Subsequently, CD154

⁺

cells were separated using the CD154 MicroBead Kit. Cells were fluorescently stained with Anti-Biotin-PE (# 130-090-756) and CD4 (VIT4)-APC (# 130-092-374) to detect CD154

⁺

cells. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.

Before separation

CMV pp65–stimulated sample	Unstimulated control
View details CD154 MicroBead Kit, human Figure 1 CD154 ⁺ T cells were isolated from human PBMCs of a CMV ⁺ donor using the CD154 MicroBead Kit, two MS Columns, and a MiniMACS™ Separator. PBMCs were stimulated for 16 hours with PepTivator CMV pp65 (# 130-093-438), a CD40 blocking-antibody was added during the stimulation to prevent down-regulation of CD154. Subsequently, CD154 ⁺ cells were separated using the CD154 MicroBead Kit. Cells were fluorescently stained with Anti-Biotin-PE (# 130-090-756) and CD4 (VIT4)-APC (# 130-092-374) to detect CD154 ⁺ cells. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.	View details CD154 MicroBead Kit, human Figure 1 CD154 ⁺ T cells were isolated from human PBMCs of a CMV ⁺ donor using the CD154 MicroBead Kit, two MS Columns, and a MiniMACS™ Separator. PBMCs were stimulated for 16 hours with PepTivator CMV pp65 (# 130-093-438), a CD40 blocking-antibody was added during the stimulation to prevent down-regulation of CD154. Subsequently, CD154 ⁺ cells were separated using the CD154 MicroBead Kit. Cells were fluorescently stained with Anti-Biotin-PE (# 130-090-756) and CD4 (VIT4)-APC (# 130-092-374) to detect CD154 ⁺ cells. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.
After separation

CMV pp65–stimulated sample

Unstimulated control

Figure 1

CD154

⁺

T cells were isolated from human PBMCs of a CMV

⁺

donor using the CD154 MicroBead Kit, two MS Columns, and a MiniMACS™ Separator. PBMCs were stimulated for 16 hours with PepTivator CMV pp65 (# 130-093-438), a CD40 blocking-antibody was added during the stimulation to prevent down-regulation of CD154. Subsequently, CD154

⁺

cells were separated using the CD154 MicroBead Kit. Cells were fluorescently stained with Anti-Biotin-PE (# 130-090-756) and CD4 (VIT4)-APC (# 130-092-374) to detect CD154

⁺

cells. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.

Figure 1

CD154

⁺

T cells were isolated from human PBMCs of a CMV

⁺

donor using the CD154 MicroBead Kit, two MS Columns, and a MiniMACS™ Separator. PBMCs were stimulated for 16 hours with PepTivator CMV pp65 (# 130-093-438), a CD40 blocking-antibody was added during the stimulation to prevent down-regulation of CD154. Subsequently, CD154

⁺

cells were separated using the CD154 MicroBead Kit. Cells were fluorescently stained with Anti-Biotin-PE (# 130-090-756) and CD4 (VIT4)-APC (# 130-092-374) to detect CD154

⁺

cells. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.

Specifications for CD154 MicroBead Kit, human

Overview

The CD154 MicroBead Kit has been developed for positive selection of activated CD154

⁺

antigen-specific CD4

⁺

T cells from human PBMCs.

Detailed product information

Background information

The CD154 antibody specifically recognizes the human CD154 antigen, a 39 kDa transmembrane glycoprotein also known as CD40L, gp39, T-BAM, TRAP, or Ly-62. CD154 is transiently up-regulated on activated CD4

⁺

T cells and plays an important role as a costimulatory molecule in T cell/antigen-presenting cell interactions through ligation of CD40. Due to its transient expression within hours after activation, CD154 can be used as a marker for activated antigen-specific CD4

⁺

T cells

¹

. Adding a CD40 blocking-antibody to the cell suspension during stimulation prevents down-regulation of CD154 expression induced by interaction with CD40 expressed on antigen-presenting cells.

Detailed separation procedure

CD154

⁺

T cells are indirectly magnetically labeled with CD154-Biotin antibody and Anti-Biotin MicroBeads UltraPure.

Columns

MS, LS, or autoMACS

^®

Columns.

Resources for CD154 MicroBead Kit, human

Documents and Protocols

Scientific posters

Rapid antigen-reactive T cell enrichment procedure enables direct ex vivo BKV-specific CD4⁺ T cell characterization

App notes/customer reports

Rapid antigen-reactive T cell enrichment (Rapid ARTE) - Workflow protocol

App notes/customer reports

Detection, enrichment, and direct phenotypic and functional characterization of antigen-specific naive and memory T cell subsets.

References for CD154 MicroBead Kit, human

Publications

Frentsch, M. et al. (2005)
Direct access to CD4
⁺
T cells specific for defined antigens according to CD154 expression.
Nat Med 11: 1118-1124
Bacher, P. et al. (2014) Antigen-specific expansion of human regulatory T cells as a major tolerance mechanism against mucosal fungi. Mucosal Immunol 7(4): 916-928
Bacher, P. et al. (2013) Antigen-reactive T cell enrichment for direct, high-resolution analysis of the human naive and memory Tʜ cell repertoire. J Immunol 190(8): 3967-3976
Schmied, S. et al. (2015)
Analysis of the functional WT1-specific T-cell repertoire in healthy donors reveals a discrepancy between CD4
⁺
and CD8
⁺
memory formation.
Immunology 145(4): 558-569

Data and images