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Authors: Kelsey Miller
Group: BioLegend
Summary: Anti-Neu5Gc Antibody Kit Protocols for Flow Cytometry, ELISA, and Western blot.

Proper citation: Kelsey Miller 2016. Anti-Neu5Gc Antibody Kit Protocols. protocols.io https://dx.doi.org/10.17504/protocols.io.e25bgg6 Copy   

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Authors: BioLegend, Inc.
Group: BioLegend

Proper citation: BioLegend, Inc. 2016. Intracellular Flow Cytometry Staining Protocol: For the Detection of Intracellular Cytokines and Other Intracellular Targets. protocols.io https://dx.doi.org/10.17504/protocols.io.ez2bf8e Copy   

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Authors: Sam Li
Group: BioLegend
Summary: Product description and procedure summary:Target cells are 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. 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 protocol has been optimized to remove washing steps after antibody cocktail and nanobeads incubations, resulting in a shorter and more convenient protocol. 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™ Isolation Kits Protocol - 1. protocols.io https://dx.doi.org/10.17504/protocols.io.7x6hpre Copy   

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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.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™ Selection Kits Protocol - 4. protocols.io https://dx.doi.org/10.17504/protocols.io.7yyhpxw Copy   

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Authors: Kelsey Miller
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 manufacturerNote: 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.

Proper citation: Kelsey Miller 2016. MojoSort™ Negative Selection Columns Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.e3mbgk6 Copy   

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Authors: Kelsey Miller
Group: BioLegend
Summary: The HLA class I ELISA is an enzyme immunoassay based on the detection of β2-microglobulin subunit of HLA class I complexes, after capturing the complex through the conjugated biotin. To this end, biotinylated HLA class I complex is first captured in streptavidin coated microtiter wells. Subsequently, HRP-conjugated anti-human β2-microglobulin is added to detect intact HLA class I complexes. Only intact HLA class I complexes are recognized. Peptides with high affinity binding will be clearly detected by this ELISA technique, while peptides with a moderate to low binding affinity for HLA class I provide a moderate to non-detectable signal. This protocol is designed to evaluate the efficiency of peptide exchange when using the Flex-T™ system.

Proper citation: Kelsey Miller 2018. Flex-T™ HLA Class I ELISA Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.tkaekse Copy   

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Authors: Kelsey Miller
Group: BioLegend

Proper citation: Kelsey Miller 2018. Immunocytochemistry Staining for Methanol Fixed Cells. protocols.io https://dx.doi.org/10.17504/protocols.io.tdhei36 Copy   

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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 1 - CD45 less than 50%. protocols.io https://dx.doi.org/10.17504/protocols.io.7wphpdn Copy   

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Authors: Sam Li
Group: BioLegend
Summary: Product description and procedure summary:Target cells are depleted by incubating your 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. 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™ Isolation Kits Protocol - 3. protocols.io https://dx.doi.org/10.17504/protocols.io.7x7hprn Copy   

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Authors: Ken Lau
Group: BioLegend

Proper citation: Ken Lau 2021. MojoSort™ Human Pan DC Isolation Kit Protocol. protocols.io https:// Copy   

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Authors: Sam Li
Group: BioLegend
Summary: Product description and procedure summary: This kit is designed for the isolation of untouched pan monocytes from PBMC. Target cells are depleted by incubating your 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.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 107cells. 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.Sample Preparation: It is strongly recommended that platelets be removed prior to the isolation of monocytes using a suitable method. See recommended platelet removal protocol below

Proper citation: Sam Li 2019. MojoSort™ Human Pan Monocyte Isolation Kit Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.7ywhpxe Copy   

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Authors: Sam Li
Group: BioLegend

Proper citation: Sam Li 2019. T Cell Activation with anti-CD3 Antibodies Protocol - Mouse. protocols.io https://dx.doi.org/10.17504/protocols.io.794hr8w Copy   

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Authors: Kelsey Miller
Group: BioLegend
Summary:  This procedure is optimized for the isolation of 107 to 1 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.Product description and procedure 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.fxnbpme Copy   

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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 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™ Human CD14+ Monocyte Isolation Kit Column Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.7bghijw Copy   

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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 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™ Isolation Kits Column Protocol - 1. protocols.io https://dx.doi.org/10.17504/protocols.io.7bahiie Copy   

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Authors: Kelsey Miller
Group: BioLegend
Summary: Note: If the percentage of CD45+ cells in your sample is less than 50%, please follow Protocol A. If it is higher than 50% then please follow protocol B.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.

Proper citation: Kelsey Miller 2016. MojoSort™ Human CD45 Nanobeads Protocols. protocols.io https://dx.doi.org/10.17504/protocols.io.e2ybgfw Copy   

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Authors: Kelsey Miller
Group: BioLegend
Summary: Using UV-induced peptide exchange, MHC/peptide monomers can be generated with conditional Flex-T™ monomers that harbor peptides of interest in their binding grooves. These new MHC monomers are subsequently multimerized using streptavidin-fluorophore conjugates. The resulting Flex-T™ reagents can be used for staining antigen-specific T cells and flow cytometric analysis. In humans, the MHC molecules are called HLA (Human Leukocyte Antigen).

Proper citation: Kelsey Miller 2017. Flex-T™ Tetramer and Cell Staining Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.hv6b69e Copy   

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Authors: Ken Lau
Group: BioLegend

Proper citation: Ken Lau 2021. MojoSort™ Whole Blood Human Neutrophil Isolation Kit Column Protocol. protocols.io https:// Copy   

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Authors: Kelsey Miller
Group: BioLegend

Proper citation: Kelsey Miller 2016. Immunohistochemistry Protocol for Keratin Antibodies. protocols.io https://dx.doi.org/10.17504/protocols.io.e2hbgb6 Copy   

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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 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™ Human Pan Monocyte Isolation Kit Column Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.7tthnnn Copy   

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