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Authors: Sam Li
Group: BioLegend
Proper citation: Sam Li 2019. Intracellular Staining With True-Phos™ Perm Buffer in Whole Blood. protocols.io https://dx.doi.org/10.17504/protocols.io.bac6iaze Copy
Authors: Sam Li
Group: BioLegend
Proper citation: Sam Li 2019. Immunohistochemistry Protocol for Beta Amyloid Products using USA Detection Kit. protocols.io https://dx.doi.org/10.17504/protocols.io.8x4hxqw Copy
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 14mL tubes and Magnet, and place the tube in the magnet for 10 minutes.Sample Preparation: Dead cell removal prior to cell isolation is recommended to achieve the highest purity and yield of mouse NK cells. Dead cells can be removed using appropriate centrifugation media or by other methods.
Proper citation: Sam Li 2019. MojoSort™ Mouse NK Cell Isolation Kit Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.7yshpwe 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 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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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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