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Authors: Kelsey Knight
Group: BioLegend

Proper citation: Kelsey Knight 2018. Th1 Polarization of Mouse CD4+ Cells. protocols.io https://dx.doi.org/10.17504/protocols.io.tncemaw Copy   

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Authors: BioLegend, Inc.
Group: BioLegend

Proper citation: BioLegend, Inc. 2017. Human T Cell Activation with anti-CD3 Anitbodies (clone UCHT1, OKT3 or HIT3a). protocols.io https://dx.doi.org/10.17504/protocols.io.hv7b69n Copy   

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Authors: BioLegend, Inc.
Group: BioLegend

Proper citation: BioLegend, Inc. 2016. Immunofluorescent Staining of Whole Blood. protocols.io https://dx.doi.org/10.17504/protocols.io.exqbfmw Copy   

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Authors: Kelsey Miller
Group: BioLegend
Summary: This protocol has been optimized to remove washing steps after antibody and nanobeads incubation, 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 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:This kit is designed for the sequential positive selection of CD4+ T cells from human peripheral blood mononuclear cells (PBMCs). Human monocytes express both CD14 and CD4. When using only CD4 Nanobeads for positive selection of human CD4 T cells, monocytes could be isolated along with the T cells. If this monocyte fraction does not impact your application, there is no need to address it. However, not including this population may be required. Thus, the first step in this kit is the depletion of CD14+ cells using a combination of biotin anti-human CD14 and Streptavidin Nanobeads. The second step is the positive selection of the CD4 T cells using directly conjugated CD4 Nanobeads. After collection of the targeted cells, downstream applications include functional assays, gene expression, phenotypic characterization, etc.

Proper citation: Kelsey Miller 2016. MojoSort™ Human CD4 T Cell Selection Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.e3gbgjw Copy   

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Authors: BioLegend, Inc.
Group: BioLegend
Summary: This is part of BioLegend's, "Intracellular Flow Cytometry Staining Protocol: For the Detection of Intracellular Cytokines and Other Intracellular Targets".

Proper citation: BioLegend, Inc. 2016. Activation and Intracellular Staining of Whole Blood: For the Detection of Intracellular Cytokines and Other Intracellular Targets. protocols.io https://dx.doi.org/10.17504/protocols.io.ez3bf8n Copy   

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Authors: BioLegend, Inc.
Group: BioLegend

Proper citation: BioLegend, Inc. 2016. Direct-Blot™ Western Blotting Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.e2gbgbw Copy   

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Authors: BioLegend, Inc.
Group: BioLegend

Proper citation: BioLegend, Inc. 2016. True-Nuclear™ Transcription Factor Staining Protocol for 5 mL Tubes. protocols.io https://dx.doi.org/10.17504/protocols.io.gq4bvyw Copy   

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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 describes surface protein staining with TotalSeq™–B and TotalSeq™–C antibodies and/or hashtag antibodies, to enable protein detection in addition to Single Cell 3’ v3 and Single Cell V(D)J Feature Barcoding technology from 10x Genomics.Please read the entire protocol before starting the experiments.

Proper citation: Sam Li 2019. TotalSeq™-B or -C with 10x Feature Barcoding Technology. protocols.io https://dx.doi.org/10.17504/protocols.io.8v5hw86 Copy   

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Authors: Kelsey Miller
Group: BioLegend
Summary: ACUITYAdvanced Biotin Free HRP Polymer Detection System for Immunohistochemistry

Proper citation: Kelsey Miller 2017. ACUITYAdvanced Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.hvjb64n Copy   

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Authors: Kelsey Miller
Group: BioLegend

Proper citation: Kelsey Miller 2017. ELISPOT Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.hv2b68e Copy   

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Authors: Sam Li
Group: BioLegend

Proper citation: Sam Li 2019. Th9 Polarization of Mouse CD4+ Cells. protocols.io https://dx.doi.org/10.17504/protocols.io.798hr9w Copy   

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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 is 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.Please read the entire protocol before starting the experiments.

Proper citation: Sam Li 2019. TotalSeq™-A Antibodies Protocol with 10x Single Cell 3' Reagent Kit v2. protocols.io https://dx.doi.org/10.17504/protocols.io.8vyhw7w 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 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™ Selection Kits Column Protocol - 4. protocols.io https://dx.doi.org/10.17504/protocols.io.7twhnpe Copy   

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Authors: Sam Li
Group: BioLegend
Summary: This protocol is essentially the same as a standard ELISA protocol, with one major exception - all animal compounds must be avoided in buffers, etc. because they contain Neu5Gc, which will interfere with antibody activity and result in misleading data. Essentially, the glycoconjugate to be evaluated is immobilized on an ELISA plate and exposed to a primary antibody. The antibody provided in this kit has been shown to identify as little as 5pmol of Neu5Gc per microgram glycoprotein, which is at or below the current detection limit for conventional DMB HPLC analysis. An advantage of the ELISA procedure is that it confirms the presence of Neu5Gc on the glycoconjugate of interest, but use caution, as it also reacts with solution/media that contains glycoconjugate.

Proper citation: Sam Li 2019. Anti-Neu5Gc Antibody Kit Protocol - ELISA. protocols.io https://dx.doi.org/10.17504/protocols.io.98sh9we Copy   

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Authors: Sam Li
Group: BioLegend

Proper citation: Sam Li 2019. Th1 Polarization of Mouse CD4+ Cells. protocols.io https://dx.doi.org/10.17504/protocols.io.795hr86 Copy   

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Authors: Sam Li
Group: BioLegend

Proper citation: Sam Li 2019. Immunocytochemistry Staining for Methanol Fixed Cells. protocols.io https://dx.doi.org/10.17504/protocols.io.97ah9ie Copy   

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Authors: Kelsey Miller
Group: BioLegend
Summary: Anti-BrdU Staining Protocol using DNAse with surface and fluorescent proteins

Proper citation: Kelsey Miller 2016. Anti-BrdU Staining Protocols Using DNAse with Surface and Fluorescent Proteins. protocols.io https://dx.doi.org/10.17504/protocols.io.e2nbgde 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 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™ Selection Kits Column Protocol - 1. protocols.io https://dx.doi.org/10.17504/protocols.io.7tuhnnw Copy   

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Authors: Sam Li
Group: BioLegend
Summary: The kit is designed for the isolation of human memory CD8 T cells from PBMC. Target 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: Sam Li 2017. MojoSort™ Human CD8 Memory T Cells Isolation Kit Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.ihjcb4n Copy   

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Authors: Kelsey Knight
Group: BioLegend

Proper citation: Kelsey Knight 2018. True-Nuclear™ Transcription Factor Staining Protocol for 96-Well U-Bottom Plate. protocols.io https://dx.doi.org/10.17504/protocols.io.tq3emyn Copy   

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