Microbeads
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.
Quantity
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.