CD4
⁺
CD25
⁺
Regulatory T Cell Isolation Kit
, human

Name: CD4+CD25+ Regulatory T Cell Isolation Kit, human
Availability: InStock

CD4
⁺
CD25
⁺
Regulatory T Cell Isolation Kit
, human

The CD4

⁺

CD25

⁺

Regulatory T Cell Isolation Kit was developed for the isolation of CD4

⁺

CD25

⁺

cells from human PBMCs. The isolation is performed in a two-step procedure.

Product data and images for CD4
⁺
CD25
⁺
Regulatory T Cell Isolation Kit
, human

Figures

Figure 1

CD4

⁺

CD25

⁺

regulatory T cells were isolated from human PBMCs. The cells were fluorescently stained as indicated and analyzed by flow cytometry using the MACSQuant® Analyzer. Gating was performed according to CD4 expression except for the CD25-APC vs. CD4 (VIT4)-VioBlue dot plot.

PBMCs before separation

View details CD4 ⁺ CD25 ⁺ Regulatory T Cell Isolation Kit , human Figure 1 CD4 ⁺ CD25 ⁺ regulatory T cells were isolated from human PBMCs. The cells were fluorescently stained as indicated and analyzed by flow cytometry using the MACSQuant® Analyzer. Gating was performed according to CD4 expression except for the CD25-APC vs. CD4 (VIT4)-VioBlue dot plot.	View details CD4 ⁺ CD25 ⁺ Regulatory T Cell Isolation Kit , human Figure 1 CD4 ⁺ CD25 ⁺ regulatory T cells were isolated from human PBMCs. The cells were fluorescently stained as indicated and analyzed by flow cytometry using the MACSQuant® Analyzer. Gating was performed according to CD4 expression except for the CD25-APC vs. CD4 (VIT4)-VioBlue dot plot.
Pre-enriched CD4⁺ cells

View details CD4 ⁺ CD25 ⁺ Regulatory T Cell Isolation Kit , human Figure 1 CD4 ⁺ CD25 ⁺ regulatory T cells were isolated from human PBMCs. The cells were fluorescently stained as indicated and analyzed by flow cytometry using the MACSQuant® Analyzer. Gating was performed according to CD4 expression except for the CD25-APC vs. CD4 (VIT4)-VioBlue dot plot.
Isolated CD4⁺CD25⁺ regulatory T cells

View details CD4 ⁺ CD25 ⁺ Regulatory T Cell Isolation Kit , human Figure 1 CD4 ⁺ CD25 ⁺ regulatory T cells were isolated from human PBMCs. The cells were fluorescently stained as indicated and analyzed by flow cytometry using the MACSQuant® Analyzer. Gating was performed according to CD4 expression except for the CD25-APC vs. CD4 (VIT4)-VioBlue dot plot.	View details CD4 ⁺ CD25 ⁺ Regulatory T Cell Isolation Kit , human Figure 1 CD4 ⁺ CD25 ⁺ regulatory T cells were isolated from human PBMCs. The cells were fluorescently stained as indicated and analyzed by flow cytometry using the MACSQuant® Analyzer. Gating was performed according to CD4 expression except for the CD25-APC vs. CD4 (VIT4)-VioBlue dot plot.
View details CD4 ⁺ CD25 ⁺ Regulatory T Cell Isolation Kit , human Figure 1 CD4 ⁺ CD25 ⁺ regulatory T cells were isolated from human PBMCs. The cells were fluorescently stained as indicated and analyzed by flow cytometry using the MACSQuant® Analyzer. Gating was performed according to CD4 expression except for the CD25-APC vs. CD4 (VIT4)-VioBlue dot plot.

Specifications for CD4
⁺
CD25
⁺
Regulatory T Cell Isolation Kit
, human

Overview

The CD4

⁺

CD25

⁺

Regulatory T Cell Isolation Kit was developed for the isolation of CD4

⁺

CD25

⁺

cells from human PBMCs. The isolation is performed in a two-step procedure.

Detailed product information

Background information

Regulatory CD4

⁺

CD25

⁺

T cells are suppressor cells that neutralize other immune cells by various mechanisms. Their characteristic marker is the transcription factor FoxP3. Regulatory CD4

⁺

CD25

⁺

T cells seem to suppress harmful immunological reactions to self or foreign antigens.

Detailed separation procedure

The isolation is performed in a two-step procedure. First, non-CD4

⁺

T cells are indirectly magnetically labeled with a cocktail of biotin-conjugated antibodies against CD8, CD14, CD15, CD16, CD19, CD36, CD56, CD123, TCRγ/δ and CD235a (glycophorin A) and Anti-Biotin MicroBeads. The labeled cells are subsequently depleted over a MACS® Column. In the second step, the flow-through fraction of pre-enriched CD4

⁺

T cells is labeled with CD25 MicroBeads for subsequent positive selection of CD4

⁺

CD25

⁺

regulatory T cells.

Downstream applications

The CD4

⁺

CD25

⁺

Regulatory T Cell Isolation Kit has been used to investigate the role of CD4

⁺

CD25

⁺

regulatory T cells in autoimmune diseases

^1,2

, antitumor immunity

^3–6

, allergy

⁷

, and in infectious diseases

⁸

. For further characterization, CD4

⁺

CD25

⁺

regulatory T cells were isolated from patients suffering from rheumatoid arthritis

¹

or multiple sclerosis

²

, from acute T cell lymphoma patients

⁴

, from tumor biopsy specimens of non-Hodgkin lymphoma patients

⁵

, after dendritic cell vaccination

⁹

as well as from patients with atopic dermatitis

⁷

.

Columns

For the first magnetic separation (depletion): LD or autoMACS

^®

Columns. For the second magnetic separation (positive selection): MS or autoMACS Columns.

Resources for CD4
⁺
CD25
⁺
Regulatory T Cell Isolation Kit
, human

Documents and Protocols

Brochures/posters

Regulatory T cell research

App notes/customer reports

Human CD4+CD25+ regulatory T cell isolation, in vitro suppression assay and analysis - Workflow protocol

App notes/customer reports

Human CD4+CD25+ regulatory T cell isolation, in vitro expansion and analysis - Workflow protocol

References for CD4
⁺
CD25
⁺
Regulatory T Cell Isolation Kit
, human

Publications

Ehrenstein, M. R. et al. (2004) Compromised function of regulatory T cells in rheumatoid arthritis and reversal by anti-TNFalpha therapy. J. Exp. Med. 200: 277-285
Hong, J. et al. (2006) CD4+ regulatory T cell responses induced by T cell vaccination in patients with multiple sclerosis. Proc. Natl. Acad. Sci. U.S.A. 103: 5024-5029
Nishikawa, H. et al. (2006)
Influence of CD4
⁺
CD25
⁺
regulatory T cells on low/high-avidity CD4
⁺
T cells following peptide vaccination.
J. Immunol. 176: 6340-6346
Karube et al. (2004) Expression of FoxP3, a key molecule in CD4CD25 regulatory T cells, in adult T-cell leukaemia/lymphoma cells. Br J Haematol 126: 81-84
Yang et al. (2006)
Intratumoral CD4
⁺
CD25
⁺
regulatory T-cell-mediated suppression of infiltrating CD4
⁺
T cells in B-cell non-Hodgkin lymphoma.
Blood 107: 3639-3646
Mougiakakos, D. et al. (2009) Naturally occurring regulatory T cells show reduced sensitivity toward oxidative stress-induced cell death. Blood 113: 3542-3545
Ou, L. S. et al. (2004) T regulatory cells in atopic dermatitis and subversion of their activity by superantigens. J. Allergy Clin. Immunol. 113: 756-763
Xu, D. et al. (2006)
Circulating and liver resident CD4
⁺
CD25
⁺
regulatory T cells actively influence the antiviral immune response and disease progression in patients with hepatitis B.
J. Immunol. 177: 739-747

Data and images