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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 781 CHF
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Description

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.

MojoSort™ reagents are also compatible with column-based cell separation systems available from other vendors. Optimized protocols for cell separation using columns from in-house testing are provided for each kit under the “Related Protocols” section, as well as representative data on the product webpage (where available). Data generated using column separators are indicated on the figure legend.

Due to the property of the beads, MojoSort™ reagents typically require dilution for optimal use on column separators. Where available, recommended dilution factors for each kit component based on in-house testing are provided under the “Application Notes” section of the webpage.

Product Details
Technical Data Sheet (pdf)

Kit Contents

Kit Contents
  • 1 mL Biotin antibody cocktail
  • 1 mL Streptavidin nanobeads
  • 100 µL APC anti-mouse CD25+ antibody
  • 100 µL anti-APC nanobeads

Product Details

Verified 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.

Product Citations
  1. Schwarz A, et al. 2023. Front Immunol. 14:1038689. PubMed

Related FAQs

Is there a way to detach your magnetic particles from the cell surface?

MojoSort™ Nanobeads: These magnetic particles cannot be removed from cells. This includes standalone Nanobeads and MojoSort kits using Nanobeads. We have found that cells are functional without the need to detach the magnetic Nanobeads.

 

MojoSort Microbeads: These magnetic particles may be released following the detailed product-specific protocol.

What is the size of your magnetic particles?

MojoSort Nanobeads: the average diameter is approximately 130 nm.

 

MojoSort Microbeads: the average diameter is approximately 1 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 product-specific information and guidance. We do not recommend using MojoSort™ particles for BD’s IMag™ or Life Technologies’ DynaMag™.

What antibodies are present in the depletion cocktails provided for isolation kits?

Please contact our technical service team for further assistance.

Go To Top Version: 3    Revision Date: 01.26.2022

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

BioLegend, the BioLegend logo, and all other trademarks are property of BioLegend, Inc. or their respective owners, and all rights are reserved.

 

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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