YILIMART - CD4 (L3T4) MicroBeads, mouse

Mouse CD4 (L3T4) MicroBeads have been developed for positive selection or depletion of mouse CD4

⁺

T cells from single-cell suspensions of lymphoid and non-lymphoid tissues or from peripheral blood.

Background information

The CD4 antigen is expressed on most thymocytes and mature T helper cells, on a subset of NKT cells and at lower levels on a subpopulation of dendritic cells. CD4 acts as an accessory molecule in the recognition of MHC class II/peptide complexes by the TCR heterodimer on CD4

⁺

T helper cells. The CD4 antigen is expressed on approximately 90% of thymocytes, 25% of splenocytes, and 55% of lymph node cells.

Downstream applications

CD4 (L3T4) MicroBeads, mouse have been designed for the isolation of mouse CD4

⁺

T cells from various tissue, such as spleen and lymph nodes, peripheral blood, brain, bone marrow, or bronchoalveolar lavage for the subsequent functional and phenotypical analysis of T helper cells

in vitro

and

in vivo

.

Furthermore, CD4 (L3T4) MicroBeads, mouse can also be used for the isolation of CD4

^–

CD8

⁺

thymocytes by depleting CD4

⁺

cells.

Examples for downstream applications include:

In vitro activation and expansion of CD4⁺ T cells (e.g. with the T Cell Activation/Expansion Kit, mouse)
Antigen-specific stimulation of CD4⁺ T cells (e.g. with PepTivator^® Peptide Pools)
Analysis of cytokine secretion and surface marker expression (e.g. with REAfinity™ Recombinant Antibodies)
CD4⁺ T cell polarization towards Tʜ cell subtypes (e.g. Tʜ1, Tʜ2, and Tʜ17 cells using the Tʜ1, Tʜ2, or Tʜ17 CytoBox).
Adoptive T cell transfer experiments (e.g. for transfer into immunodeficient, knock-out, parasite infected, and tumor-bearing mice or to passively transfer a disease with auto-antigen-stimulated CD4⁺ T cells in an EAE mouse model).
Examination of the regulatory role of cytokines in autoimmune disease and infection.
Studies on the function of costimulatory molecules (e.g. OX40-OX40 ligand interactions).
CD4⁺ T cell isolation from CNS suspensions of MHV-infected mice for evaluation of their chemokine expression pattern.

Columns

For positive selection: MS, LS, or autoMACS

^®

Columns. For depletion: LD or autoMACS Columns.

Figure 1

A single-cell suspension from mouse spleen was prepared using the gentleMACS™ Dissociator. CD4

⁺

T cells were isolated from this single-cell suspension using the CD4 (L3T4) MicroBeads, an LS Column, and a QuadroMACS™ Separator. Cells were fluorescently stained with REAfinity™ Antibodies CD45-VioGreen™, CD4-VioBlue

^®

and CD8a-PE-Vio

^®

770 and analyzed by flow cytometry using the MACSQuant

^®

Analyzer. Viable leukocytes were gated for analysis based on scatter signals, 7-AAD Staining Solution fluorescence, and CD45 expression.