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Non CD3+ T cells are depleted by incubating your sample with the biotin antibody cocktail followed by incubation with magnetic Streptavidin Nanobeads. The magnetically labeled fraction is retained by the use of a magnetic separator. The untouched CD3+ T cells are collected by decanting the liquid in a clean tube. These are your cells of interest; do not discard the liquid. Some of the downstream applications include functional assays, gene expression, phenotypic characterization, etc.
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.
- Kit Contents
For Cat# 480021:
- 100 µl Biotin-Antibody Cocktail
- 100 µl Streptavidin Nanobeads
For Cat# 480022
- 1 ml Biotin-Antibody Cocktail
- 1 ml Streptavidin Nanobeads
- Verified Reactivity
Cocktail: Phosphate-buffered solution containing 0.09% sodium azide, pH 7.2.
Streptavidin Nanobeads: Aqueous solution containing BSA and 0.05% sodium azide.
The antibodies were purified by affinity chromatography, and conjugated with biotin under optimal conditions.
Streptavidin Nanobeads: Streptavidin-coated magnetic beads.
- Storage & Handling
- Antibody cocktail and Streptavidin Nanobeads should be stored undiluted between 2°C and 8°C.
Cell Separation (MojoSort™) - Quality tested
- Recommended Usage
10 µl of antibody cocktail for 1x107 cells in 100 µl of buffer.
10 µl Streptavidin Nanobeads for 1x107 cells in 100 µl of buffer.
For this kit, incubation with antibody cocktail and Streptavidin Nanoparticles can be done for 5 to 15 minutes, without loss in purity and yield.
- Application Notes
The expected purity is >95% CD3+ cells post-separation of human peripheral blood mononuclear cells from healthy donors, analyzed by flow cytometry
This kit is designed for the isolation of untouched CD3+ T cells from peripheral blood mononuclear cells (PBMCs).
Each lot has been individually optimized. Do not mix and match components from different lots or different kits.
Antibody or cocktail dilution to use in column: 2.5X
Nanobead dilution to use in columns: 4X
- Additional Product Notes
- Product Citations
- Biology Area
- Molecular Family
- CD Molecules, TCRs
- Gene ID
- View information about CD3 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.
- 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™.
- What antibodies are present in the depletion cocktails provided for isolation kits?
Please contact our technical service team for further assistance.