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Mouse CD45.1+ cells are either selected or depleted by incubating the sample with a biotin conjugated anti-mouse CD45.1 antibody and then magnetic Streptavidin Nanobeads. The magnetically labeled fraction is retained by the use of a magnetic separator. After collection of the CD45.1+ 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. No. 480117:
- 1 vial containing 200 µl of Biotin anti-mouse CD45.1 antibody (clone A20)
- 1 vial containing 200 µl Streptavidin Nanobeads
For Cat. No. 480118:
- 2 vials containing 1 ml each of Biotin anti-mouse CD45.1 antibody (clone A20)
- 2 vials containing 1 ml each of Streptavidin Nanobeads
- Verified Reactivity
Antibody: Phosphate buffer solution containing 0.09% sodium azide, pH 7.2 with stabilizer.
Streptavidin Nanobeads: Aqueous solution containing 0.05% sodium azide and 0.3% BSA.
The antibodies were purified by affinity chromatography, and conjugated wtih biotin under optimal conditions.
Streptavidin nanobeads: Protein-coated magnetic beads.
- Storage & Handling
- All components should be stored undiluted between 2°C and 8°C.
Cell Separation (MojoSort™) - Quality tested
- Recommended Usage
Volume of Streptavidin Nanobeads should be adjusted depending on the starting percentage of CD45.1+ cells to be isolated. Use the table below as an example when working with a mixture of SJL (CD45.1+) and C57BL/6 (CD45.2+) mouse splenocytes. For 1x107 cells in 100 µl of buffer, use the following volumes:
SJL + C57BL/6 Mixture (spleen)
Approximate Starting CD45.1 Frequency
Optimal Nanobeads Volume
10% SJL + 90% C57BL/6
25% SJL + 75% C57BL/6
50% SJL + 50% C57BL/6
75% SJL + 25% C57BL/6
The volumes indicated in the table are for the use of MojoSort™ magnet (Cat. Nos. 480019/480020). For low frequency cells, pre-dilute the Streptavidin Nanobeads in order to pipette a minimum of 5 µl of any solution. For example, to isolate CD45.1+ cells from a mixture of 25% SJL and 75% C57BL/6, pre-dilute 10 µl of Streptavidin Nanobeads in 40 µl of MojoSort™ buffer (Cat. No. 480017) and add 10 µl of that dilution per sample. Avoid working with small volumes.
- Application Notes
This kit is designed for the positive selection or depletion of mouse CD45.1+ cells from lymphoid tissue.
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: 15X*
Nanobead dilution to use in columns: 4X*
*Note: Suggested dilutions for column use above determined based on a starting cell population consisting of a 1:1 mix of CD45.1+ and CD45.2+ cells. Further optimization may be necessary for different degrees of heterogeneity in starting cell populations.
- Biology Area
- Cell Biology, Immunology, Inhibitory Molecules, Neuroscience, Neuroscience Cell Markers
- Molecular Family
- CD Molecules
- Gene ID
- View information about CD45.1 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.