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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 2020. MojoSort™ Whole Blood Human CD8 Nanobeads Whole Blood Column Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.bbujinun Copy
Authors: Sam Li
Group: BioLegend
Proper citation: Sam Li 2019. T Cell Activation with anti-CD3 Antibodies Protocol - Human. protocols.io https://dx.doi.org/10.17504/protocols.io.793hr8n 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™ Human CD14 Nanobeads Column Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.7ahhib6 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™ Nanobeads Column Protocol - 2. protocols.io https://dx.doi.org/10.17504/protocols.io.69ehh3e Copy
Authors: Sam Li
Group: BioLegend
Summary: Product description and procedure summary: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 14 mL tubes and Magnet, and place the tube in the magnet for 10 minutes.
Proper citation: Sam Li 2019. MojoSort™ Nanobeads Protocol - 2. protocols.io https://dx.doi.org/10.17504/protocols.io.7u7hnzn Copy
Authors: Sam Li
Group: BioLegend
Proper citation: Sam Li 2019. Treg Polarization of Mouse CD4+ Cells. protocols.io https://dx.doi.org/10.17504/protocols.io.792hr8e Copy
Authors: Sam Li
Group: BioLegend
Summary: Product description and procedure summary: Target cells are positively selected or depleted by incubating the sample with an anti-human PE conjugated antibody, followed by incubation with magnetic anti-PE Nanobeads. The magnetically labeled fraction is retained by the use of a magnetic separator. These are the PE+ cells, do not discard them if those are the cells of interest. Some of the 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™ Mouse anti-PE Nanobeads Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.7xshpne Copy
Authors: Sam Li
Group: BioLegend
Proper citation: Sam Li 2019. Direct-Blot™ Western Blotting Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.98rh9v6 Copy
Authors: Sam Li
Group: BioLegend
Proper citation: Sam Li 2019. ELISPOT Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.98yh9xw Copy
Authors: Sam Li
Group: BioLegend
Summary: Product description and procedure summary:The cells targeted by the Nanobeads are selected 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™ Mouse CD45 Nanobeads Protocol - Selection. protocols.io https://dx.doi.org/10.17504/protocols.io.7wihpce Copy
Authors: Sam Li
Group: BioLegend
Proper citation: Sam Li 2019. ACUITYAdvanced Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.8wbhxan Copy
Authors: Sam Li
Group: BioLegend
Proper citation: Sam Li 2019. Western Blotting Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.98mh9u6 Copy
Authors: Sam Li
Group: BioLegend
Summary: Product description and procedure summary:
If your target cells are the labeled cells (the positive fraction), use the Streptavidin Nanobeads Protocol – Positive Selection. If your target cells are the unlabeled cells (negative fraction), use the Streptavidin Nanobeads Protocol - Negative Selection.
Target cells are isolated by incubating your sample with a Biotin-conjugated antibody or 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. These are the cells of interest; do not discard the liquid. Some of the 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 14 mL tubes and Magnet, and place the tube in the magnet for 10 minutes.
Proper citation: Sam Li 2019. MojoSort™ Streptavidin Nanobeads Protocol - Negative Selection. protocols.io https://dx.doi.org/10.17504/protocols.io.7u8hnzw Copy
Authors: Sam Li
Group: BioLegend
Summary: Buyer is solely responsible for determining whether Buyer has all intellectual property rights that are necessary for Buyer's intended uses of the BioLegend TotalSeq™ products. For example, for any technology platform Buyer uses with TotalSeq™, it is Buyer's sole responsibility to determine whether it has all necessary third party intellectual property rights to use that platform and TotalSeq™ with that platform.The following protocol combines highly multiplexed protein marker detection with unbiased transcriptome profiling for thousands of single cells. Epitope detection and sample multiplexing (Cell Hashing) are enabled by BioLegend’s TotalSeq™ antibody-oligo conjugates. TotalSeq™ reagents are compatible with Cellular Indexing of Transcriptomes and Epitopes by sequencing (CITE-seq), RNA Expression And Protein sequencing assay (REAP-seq), and similar workflows.
Proper citation: Sam Li 2019. TotalSeq™-A Antibodies and Cell Hashing with 10x Single Cell 3' Reagent Kit v2. protocols.io https://dx.doi.org/10.17504/protocols.io.8v3hw8n Copy
Authors: Sam Li
Group: BioLegend
Summary: Mouse CD326 (Ep-CAM) cells are selected by incubating the sample with biotin conjugated anti-mouse CD326 antibody after blocking with TruStain FcX™ (anti-mouse CD16/32 antibody) then followed by Streptavidin Nanobeads (Cat. No. 480015/480016). The magnetically labeled fraction is retained by the use of a magnetic separator. After collection of the CD326 (Ep-CAM) expressing cells, downstream applications include functional assays, gene expression, phenotypic characterization, etc. In addition to CD326 (Ep-CAM) expression on epithelial and tumor cells, CD326 is also expressed on subtypes of leukocytes, which should be depleted using MojoSort™ Mouse CD45 Nanobeads (Cat. No. 480027/480028) before CD326 positive selection.
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™ Mouse CD326 (Ep-CAM) Positive Selection Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.72ahqae Copy
Authors: Sam Li
Group: BioLegend
Proper citation: Sam Li 2019. Anti-DYKDDDDK Tag (L5) Affinity Gel Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.98kh9uw Copy
Authors: Sam Li
Group: BioLegend
Proper citation: Sam Li 2019. Th2 Polarization of Mouse CD4+ Cells. protocols.io https://dx.doi.org/10.17504/protocols.io.796hr9e Copy
Authors: Sam Li
Group: BioLegend
Summary: Astrocytes are the major glial cells in the central nervous system and are involved in the regulation of numerous processes like energy metabolism, ion homeostasis, and synaptic plasticity. Primary astrocytes derived from rodent brains provide a rapid and easy solution for studying astrocyte biology and morphology. These cultured cells are an invaluable tool for elucidating underlying mechanisms of diseases such as: 1Astrocytes are the major glial cells in the central nervous system and are involved in the regulation of numerous processes like energy metabolism, ion homeostasis, and synaptic plasticity. Primary astrocytes derived from rodent brains provide a rapid and easy solution for studying astrocyte biology and morphology. These cultured cells are an invaluable tool for elucidating underlying mechanisms of diseases such as:StrokeEpilepsyAmyotrophic lateral sclerosisParkinson’s diseaseAlzheimer’s diseaseTraumaThis protocol demonstrates how to isolate and culture primary astrocytes from mouse pup brains. The purity and morphology of astrocytes is assessed by immunocytochemistry (ICC) using well-established astrocytic markers, including GFAP.StrokeEpilepsyAmyotrophic lateral sclerosisParkinson’s diseaseTraumaAlzheimer's disease 1Astrocytes are the major glial cells in the central nervous system and are involved in the regulation of numerous processes like energy metabolism, ion homeostasis, and synaptic plasticity. Primary astrocytes derived from rodent brains provide a rapid and easy solution for studying astrocyte biology and morphology. These cultured cells are an invaluable tool for elucidating underlying mechanisms of diseases such as:StrokeEpilepsyAmyotrophic lateral sclerosisParkinson’s diseaseAlzheimer’s diseaseTraumaThis protocol demonstrates how to isolate and culture primary astrocytes from mouse pup brains. The purity and morphology of astrocytes is assessed by immunocytochemistry (ICC) using well-established astrocytic markers, including GFAP. 1Astrocytes are the major glial cells in the central nervous system and are involved in the regulation of numerous processes like energy metabolism, ion homeostasis, and synaptic plasticity. Primary astrocytes derived from rodent brains provide a rapid and easy solution for studying astrocyte biology and morphology. These cultured cells are an invaluable tool for elucidating underlying mechanisms of diseases such as:StrokeEpilepsyAmyotrophic lateral sclerosisParkinson’s diseaseAlzheimer’s diseaseTraumaThis protocol demonstrates how to isolate and culture primary astrocytes from mouse pup brains. The purity and morphology of astrocytes is assessed by immunocytochemistry (ICC) using well-established astrocytic markers, including GFAP. 1Astrocytes are the major glial cells in the central nervous system and are involved in the regulation of numerous processes like energy metabolism, ion homeostasis, and synaptic plasticity. Primary astrocytes derived from rodent brains provide a rapid and easy solution for studying astrocyte biology and morphology. These cultured cells are an invaluable tool for elucidating underlying mechanisms of diseases such as:StrokeEpilepsyAmyotrophic lateral sclerosisParkinson’s diseaseAlzheimer’s diseaseTraumaThis protocol demonstrates how to isolate and culture primary astrocytes from mouse pup brains. The purity and morphology of astrocytes is assessed by immunocytochemistry (ICC) using well-established astrocytic markers, including GFAP. 1Astrocytes are the major glial cells in the central nervous system and are involved in the regulation of numerous processes like energy metabolism, ion homeostasis, and synaptic plasticity. Primary astrocytes derived from rodent brains provide a rapid and easy solution for studying astrocyte biology and morphology. These cultured cells are an invaluable tool for elucidating underlying mechanisms of diseases such as:StrokeEpilepsyAmyotrophic lateral sclerosisParkinson’s diseaseAlzheimer’s diseaseTraumaThis protocol demonstrates how to isolate and culture primary astrocytes from mouse pup brains. The purity and morphology of astrocytes is assessed by immunocytochemistry (ICC) using well-established astrocytic markers, including GFAP. 1Astrocytes are the major glial cells in the central nervous system and are involved in the regulation of numerous processes like energy metabolism, ion homeostasis, and synaptic plasticity. Primary astrocytes derived from rodent brains provide a rapid and easy solution for studying astrocyte biology and morphology. These cultured cells are an invaluable tool for elucidating underlying mechanisms of diseases such as:StrokeEpilepsyAmyotrophic lateral sclerosisParkinson’s diseaseAlzheimer’s diseaseTraumaThis protocol demonstrates how to isolate and culture primary astrocytes from mouse pup brains. The purity and morphology of astrocytes is assessed by immunocytochemistry (ICC) using well-established astrocytic markers, including GFAP.
Proper citation: Sam Li 2019. Isolation and Culture of Primary Mouse Astrocytes. protocols.io https://dx.doi.org/10.17504/protocols.io.97eh9je Copy
Authors: Sam Li
Group: BioLegend
Proper citation: Sam Li 2019. Ki-67 Staining Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.bac2iaye Copy
Authors: Sam Li
Group: BioLegend
Proper citation: Sam Li 2019. Immunohistochemistry Protocol for Ultra Streptavidin Detection Kits (USA). protocols.io https://dx.doi.org/10.17504/protocols.io.95rh856 Copy
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