Human CD3+ cells are either selected or depleted by incubating the sample with a biotin conjugated anti-human CD3 antibody and magnetic Streptavidin Nanobeads. The magnetically labeled fraction is retained by the use of a magnetic separator. After collection of the CD3+ expressing cells, 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# 480133:
- 1 vial containing 200 µl of Biotin anti-human CD3 antibody
- 1 vial containing 200 µl Streptavidin Nanobeads
For Cat# 480134:
- 2 vials containing 1mL each of Biotin anti-human CD3 antibody
- 2 vials containing 1mL each of Streptavidin Nanobeads
- Verified Reactivity
Cocktail: Phosphate buffer solution containing 0.09% sodium azide, 0.5% BSA, pH 7.2 with stabilizer.
Particle: Aqueous solution containing 0.05% sodium azide and 0.3% BSA.
Biotin Antibody: The antibody was purified by affinity chromatography, and conjugated with biotin under optimal conditions.
Conjugated nanobeads: Protein-coated magnetic beads.
- Recommended Usage
10 µL of Biotin anti-human CD3 antibody for 1 x 107 cells in 100 µL of buffer.
10 µL of Streptavidin Nanobeads for 1 x 107 cells in 100 µL of buffer.
- Application Notes
The expected purity is >80% CD3+/CD14- cells post-separation of human peripheral blood mononuclear cells from healthy donors, analyzed by flow cytometry
This kit is designed for the positive selection or depletion of CD3+ cells from human 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: 7X
Nanobead dilution to use in columns: 15X
- 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.