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Mouse CD4+CD25+ Regulatory T cells are isolated in a two-step separation process. The cells are first incubated with the biotin conjugated antibody cocktail, followed by the Streptavidin nanobeads, to isolate total CD4+ T cells. The second step consists of a positive selection of CD25+ cells using APC anti-mouse CD25 antibody and anti-APC nanobeads. The magnetically labeled fraction is retained by the use of a magnetic separator. After collection of the CD4+CD25+ cells, downstream applications include functional assays, gene expression, phenotypic characterization, etc. One test is equivalent to process 108 cells, approximately one mouse spleen.Product Details
- Kit Contents
- 1 vial containing 1 mL of Biotin antibody cocktail: Biotin anti- CD8a, CD11b, CD11c, CD19, CD24, CD45R/B220, CD49b, CD105, I-A/I-E (MHC II), TCR-γδ, TER-119/Erythroid
- 1 vial containing 1 mL of Streptavidin nanobeads
- 1 vial containing 100 µL of APC anti-mouse CD25+ antibody
- 1 vial containing 100 µL anti-APC nanobeads
Antibody: Phosphate buffer solution containing 0.09% sodium azide, pH 7.2
Streptavidin nanobeads: Aqueous solution containing BSA and 0.05% sodium azide
Anti-APC nanobeads: Aqueous solution containing BSA and 0.05% sodium azide
- The antibodies were purified by affinity chromatography, and conjugated with biotin or APC under optimal conditions. Conjugated nanobeads: Protein-coated magnetic beads.
- Storage & Handling
- All components should be stored undiluted between 2°C and 8°C.
- Recommended Usage
One test is equivalent to process 108 cells, approximately one mouse spleen.
Use the table below as a guideline when working with mouse splenocytes. For 108 cells in 1mL of buffer use the following volumes:
Cell number Biotin antibody cocktail Streptavidin nanobeads APC anti-mouse CD25 anti-APC nanobeads
(1 x 108 cells)
100 μL 100 μL 10 μL 10 μL
The volumes indicated in the table are optimized with the use of a MojoSort™ magnet (Cat. Nos. 480019, 480020). Scale up volumes as needed when working with larger numbers of cells.
- Application Notes
This kit is designed for the positive isolation or depletion of mouse CD4+CD25+ cells from mouse spleen.
Each lot has been individually optimized. Do not mix and match components from different lots or different kits.
- Gene ID
- View information about Regulatory T on UniProt.org
- Is there a way to detach your magnetic particles from the cell surface?
No, not currently. We have found that cells are functional without the need to detach the magnetic Nanobeads.
- What is the size of your magnetic particles?
The average diameter is approximately 130 nm.
- Can your magnetic particles be sterile filtered?
Yes, they can be sterile filtered as the particles are smaller than 0.22 µm.
- Are MojoSort™ Nanobeads compatible with other commercially available magnetic separation systems?
MojoSort™ magnetic particles can be used with other commercially available magnetic separators, both free standing magnets and column-based systems. Because MojoSort™ protocols are optimized for the MojoSort™ separator, the protocols may need to be adjusted for other systems. Please contact BioLegend Technical Service for more information and guidance. We do not recommend using MojoSort™ particles for BD’s IMag™ or Life Technologies’ DynaMag™.