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Cell populations other than total monocytes (CD14+ and CD14+CD16+ cells) are depleted by incubating the 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 monocytes are collected by decanting the liquid in a clean tube. These are the cells of interest; do not discard the liquid. Some of the downstream applications include functional assays, gene expression, phenotypic characterization, etc.
MojoSort™ Human Pan Monocyte Isolation Kit Protocol
- Kit Contents
For Cat# 480059:
• 200 µl Biotin-Antibody Cocktail
• 200 µl Streptavidin Nanobeads
• 100 µl Human TruStain FcX™ (Fc Receptor Blocking Solution)
For Cat# 480060:
• 2 vials of 1 ml Biotin-Antibody Cocktail each
• 2 vials of 1 ml Streptavidin Nanobeads each
• 1 ml Human TruStain FcX™ (Fc Receptor Blocking Solution)
- Verified Reactivity
- Cocktail: Phosphate buffer 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.Strepavidin Nanobeads: Streptavidin-coated magnetic beads.
- Storage & Handling
- Antibody cocktail, Streptavidin Nanobeads, and Human TruStain FcX™ (Fc Receptor Blocking Solution) should be stored undiluted between 2°C and 8°C.
Cell Separation (MojoSort™) - Quality tested
- Recommended Usage
10 µl of antibody cocktail 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.
5 µl of Human TruStain FcX™ (Fc Receptor Blocking Solution) for 1 x 107 cells in 100 µl of buffer.
- Application Notes
This kit is designed for the isolation of untouched pan monocytes from peripheral blood mononuclear cells (PBMCs).
Antibody or cocktail dilution to use in column: 9X
Nanobead dilution to use in columns: 2.2X
- Product Citations
- Gene ID
- 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.