MojoSort™ Mouse CD4+CD25+ Regulatory T Cell Isolation Kit

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MojoSort_Mouse_CD4-CD25_Regulatory_T_Cell_Isolation_Kit_1_062620.png
A single cell suspension from C57BL/6 mouse spleen was prepared for the negative isolation of CD4+ cells and subsequent positive selection of CD25+ cells using the MojoSort™ Mouse CD4+CD25+ Regulatory T Cell Isolation Kit. Left plot: Cells after CD4 negative isolation and CD25 positive selection. Right plot: Cells prior to CD4 depletion. Cells were fixed and permeabilized using the True-Nuclear™ Transcription Buffer Set, then intracellularly stained with anti-mouse FOXP3 Alexa Fluor® 488. Dead cells were excluded by Helix NP™ Blue.
  • MojoSort_Mouse_CD4-CD25_Regulatory_T_Cell_Isolation_Kit_1_062620.png
    A single cell suspension from C57BL/6 mouse spleen was prepared for the negative isolation of CD4+ cells and subsequent positive selection of CD25+ cells using the MojoSort™ Mouse CD4+CD25+ Regulatory T Cell Isolation Kit. Left plot: Cells after CD4 negative isolation and CD25 positive selection. Right plot: Cells prior to CD4 depletion. Cells were fixed and permeabilized using the True-Nuclear™ Transcription Buffer Set, then intracellularly stained with anti-mouse FOXP3 Alexa Fluor® 488. Dead cells were excluded by Helix NP™ Blue.
  • MojoSort_Mouse_CD4-CD25_Regulatory_T_Cell_Isolation_Kit_2_062620.png
    A single cell suspension from C57BL/6 mouse spleen was prepared for the negative isolation of CD4+ cells and subsequent positive selection of CD25+ cells using the MojoSort™ Mouse CD4+CD25+ Regulatory T Cell Isolation Kit. Left plot: Cells after CD4 negative isolation and CD25 positive selection. Right plot: Cells prior to CD4 depletion. Cells were fixed and permeabilized using the True-Nuclear™ Transcription Buffer Set, then intracellularly stained with anti-mouse FOXP3 Alexa Fluor® 488. Dead cells were excluded by Helix NP™ Blue.
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480137 10 tests 540,00€
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Description

Mouse CD4+CD25Regulatory 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
Technical Data Sheet (pdf)

Kit Contents

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

Product Details

Reactivity
Mouse
Formulation
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
Preparation
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.
Application

Cell Separation (MojoSort™) - Quality tested

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

Mouse Splenocytes

(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.

Related FAQs

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™.

Go To Top Version: 2    Revision Date: 10-20-2020

For research use only. Not for diagnostic use. Not for resale. BioLegend will not be held responsible for patent infringement or other violations that may occur with the use of our products.

 

*These products may be covered by one or more Limited Use Label Licenses (see the BioLegend Catalog or our website, www.biolegend.com/ordering#license). BioLegend products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products, reverse engineer functionally similar materials, or to provide a service to third parties without written approval of BioLegend. By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. Unless otherwise indicated, these products are for research use only and are not intended for human or animal diagnostic, therapeutic or commercial use.

 

BioLegend Inc., 8999 BioLegend Way, San Diego, CA 92121 www.biolegend.com
Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

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