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Authors: Sam Li
Group: BioLegend
Summary: Precision Count Beads™ are designed for counting the absolute number of cells in a complex mix population and other particles by flow cytometry. Precision Count Beads™ are excited by a variety of lasers including violet (405nm), blue (488nm), yellow/green (562nm), and red (633nm).
Proper citation: Sam Li 2019. Precision Count Beads™ Protocol and Applications. protocols.io https://dx.doi.org/10.17504/protocols.io.baariad6 Copy
Authors: BioLegend, Inc., Kelsey Knight
Group: BioLegend
Proper citation: BioLegend, Inc., Kelsey Knight 2018. Immunocytochemistry Staining Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.tdmei46 Copy
Authors: Sam Li
Group: BioLegend
Summary: BioLegend MojoSort™ nanobeads work in commonly used separation columns, based on our internal research as well as validation by external testing by academic labs. This simple protocol consists of following the MojoSort™ protocol to label the cells with pre-diluted MojoSort™ reagents and using the columns as indicated by the manufacturer.Note: Due to the properties of our beads, it may be possible to use far fewer beads that with other commercial suppliers. We recommend a titration to find the best dilution factor. However, as a general rule, dilutions ranging from 1:3 to 1:20 for the Nanobeads can be used. Please contact BioLegend Technical Service ([email protected]) if further assistance is needed.
Proper citation: Sam Li 2019. MojoSort™ Mouse CD4 Nanobeads Column Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.696hh9e Copy
Authors: Sam Li
Group: BioLegend
Summary: Product description and procedure summary:Target cells are either selected or depleted by incubating the sample with the biotin antibody cocktail followed by incubation with magnetic Streptavidin Nanobeads (Cat. No. 480015/480016). The magnetically labeled fraction is retained by the use of a magnetic separator. The untouched cells are collected. If these are the cells of interest; do not discard the liquid. Some of the downstream applications include functional assays, gene expression, phenotypic characterization, etc.Sample Preparation:Enzymatic digestion of mouse brain followed by myelin removal is recommended to achieve the highest purity and yield. There are several protocols published that can be applied. As a general guideline, Trypsin digestion followed by a 70/37/30% percoll gradient will increase final purity and yield.Scale up volumes if using 14 ml tubes and Magnet, and place the tube in the magnet for 10 minutes.
Proper citation: Sam Li 2019. MojoSort™ Selection Kits Protocol - 5. protocols.io https://dx.doi.org/10.17504/protocols.io.7z9hp96 Copy
Authors: Sam Li
Group: BioLegend
Proper citation: Sam Li 2019. Whole Mouse Brain Processing for Microglia Isolation, Cell Separation, and Flow Cytometry. protocols.io https://dx.doi.org/10.17504/protocols.io.97dh9i6 Copy
Authors: Kelsey Miller
Group: BioLegend
Proper citation: Kelsey Miller 2016. Intracellular Staining With True-Phos™ Perm Buffer in Whole Blood. protocols.io https://dx.doi.org/10.17504/protocols.io.e2dbga6 Copy
Authors: Ken Lau
Group: BioLegend
Proper citation: Ken Lau 2021. MojoSort™ Human CD56 Nanobeads Protocol. protocols.io https:// Copy
Authors: Sam Li
Group: BioLegend
Summary: BioLegend MojoSort™ nanobeads work in commonly used separation columns, based on our internal research as well as validation by external testing by academic labs. This simple protocol consists of following the MojoSort™ protocol to label the cells with pre-diluted MojoSort™ reagents and using the columns as indicated by the manufacturer.Note: Due to the properties of our beads, it may be possible to use far fewer beads than with other commercial suppliers. We recommend a titration to find the best dilution factor. However, as a general rule, dilutions ranging from 1:3 to 1:20 for the Nanobeads can be used. Please contact BioLegend Technical Service ([email protected]) if further assistance is needed.
Proper citation: Sam Li 2019. MojoSort™ Mouse CD11c Nanobeads Column Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.7abhian Copy
Authors: Kelsey Miller
Group: BioLegend
Summary: Cell Fixation and Permeabilization Protocol Using 70% Ethanol
Proper citation: Kelsey Miller 2016. Ethanol Fixation Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.e24bggw Copy
Authors: BioLegend, Inc.
Group: BioLegend
Proper citation: BioLegend, Inc. 2016. Treg Polarization of Mouse CD4+ Cells. protocols.io https://dx.doi.org/10.17504/protocols.io.ex7bfrn Copy
Authors: Sam Li
Group: BioLegend
Proper citation: Sam Li 2019. Intracellular Staining With True-Phos™ Perm Buffer in Cell Suspensions. protocols.io https://dx.doi.org/10.17504/protocols.io.bac4iayw Copy
Authors: Sam Li
Group: BioLegend
Summary: Product description and procedure summary:If the percentage of CD45+ cells in your sample is less than 50%, please follow Human CD45 Nanobeads Protocol 1. If it is higher than 50% then please follow Human CD45 Nanobeads Protocol 2.
The cells targeted by the Nanobeads are either selected or depleted by incubating your sample with the directly conjugated magnetic particles. The magnetically labeled fraction is retained by the use of a magnetic separator. After collection of the targeted cells, downstream applications include functional assays, gene expression, phenotypic characterization, etc.Note: This procedure is optimized for the isolation of 107 to 2 x 108 cells per tube. If working with fewer than 107 cells, keep volumes as indicated for 107 cells. For best results, optimize the conditions to your specific cell number and tissue. Prepare fresh MojoSort™ Buffer solution by diluting the 5X concentrate with sterile distilled water.Scale up volumes if using 14mL tubes and Magnet, and place the tube in the magnet for 10 minutes.
Proper citation: Sam Li 2019. MojoSort™ Human CD45 Nanobeads Protocol 2 - CD45 greater than 50%. protocols.io https://dx.doi.org/10.17504/protocols.io.7wqhpdw Copy
Authors: BioLegend, Inc.
Group: BioLegend
Proper citation: BioLegend, Inc. 2016. Th17 Polarization of Mouse CD4+ Cells. protocols.io https://dx.doi.org/10.17504/protocols.io.ezrbf56 Copy
Authors: Kelsey Miller
Group: BioLegend
Summary: Precision Count Beads™ are designed for counting the absolute number of cells in a complex mix population and other particles by flow cytometry. Precision Count Beads™ are excited by a variety of lasers including violet (405nm), blue (488nm), yellow/green (562nm), and red (633nm).
Proper citation: Kelsey Miller 2016. Precision Count Beads™ Protocol and Applications. protocols.io https://dx.doi.org/10.17504/protocols.io.e3cbgiw Copy
Authors: Kelsey Miller
Group: BioLegend
Proper citation: Kelsey Miller 2016. Ki-67 Staining Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.fabbian Copy
Authors: Kelsey Miller
Group: BioLegend
Summary: Non Neutrophil 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 cells are collected. These are the cells of interest; do not discard the liquid. Some of the downstream applications include functional assays, gene expression, phenotypic characterization, etc.
Proper citation: Kelsey Miller 2016. Mojosort™ Mouse Neutrophil Isolation Kit Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.fkabkse Copy
Authors: BioLegend, Inc.
Group: BioLegend
Proper citation: BioLegend, Inc. 2016. Human T Cell Activation with anti-CD3 Anitbodies (clone UCHT1, OKT3 or HIT3a). protocols.io https://dx.doi.org/10.17504/protocols.io.extbfnn Copy
Authors: Sam Li
Group: BioLegend
Summary: BioLegend MojoSort™ nanobeads work in commonly used separation columns, based on our internal research as well as validation by external testing by academic labs. This simple protocol consists of following the MojoSort™ protocol to label the cells with pre-diluted MojoSort™ reagents and using the columns as indicated by the manufacturer.Note: Due to the properties of our beads, it may be possible to use far fewer beads and less antibody cocktail that with other commercial suppliers. We recommend a titration to find the best dilution factor. However, as a general rule, dilutions ranging from 1:2 to 1:10 for the antibody cocktail can be used. Dilutions ranging from 1:5 to 1:20 for the Streptavidin Nanobeads can be used. Please contact BioLegend Technical Service ([email protected]) if further assistance is needed.
Proper citation: Sam Li 2019. MojoSort™ Mouse Neutrophil Isolation Kit Column Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.7bjhikn Copy
Authors: Sam Li
Group: BioLegend
Proper citation: Sam Li 2019. Anti-BrdU Staining Protocols Using DNAse with Surface and Fluorescent Proteins. protocols.io https://dx.doi.org/10.17504/protocols.io.bac9iaz6 Copy
Authors: BioLegend, Inc.
Group: BioLegend
Proper citation: BioLegend, Inc. 2016. True-Nuclear™ Transcription Factor Staining Protocol for 96-Well, U-Bottom Plate. protocols.io https://dx.doi.org/10.17504/protocols.io.gq5bvy6 Copy
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