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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: 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 2017. Flex-T™ HLA Class I ELISA Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.mcsc2we Copy
Authors: Kelsey Miller
Group: BioLegend
Summary: Protocols for Anti-BrdU Staining Using DNAse with Surface and Fluorescent Proteinsand Anti-BrdU Staining Using 70% Ethanol and 2N HCl
Proper citation: Kelsey Miller 2016. Anti-BrdU Staining Protocols. protocols.io https://dx.doi.org/10.17504/protocols.io.e23bggn Copy
Authors: Kelsey Miller
Group: BioLegend
Proper citation: Kelsey Miller 2016. Immunohistochemistry Protocol for Ultra Streptavidin Detection Kits (USA). protocols.io https://dx.doi.org/10.17504/protocols.io.e2kbgcw Copy
Authors: Kelsey Miller
Group: BioLegend
Summary: This procedure is optimized for the isolation of 106 cells per test. If working with fewer than 106
cells, keep volumes as indicated for 106 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:
Target cells are positively selected or depleted by incubating the sample with the biotin anti‐mouse CX3CR1
antibody followed by incubation with magnetic Streptavidin Nanobeads. The magnetically labeled fraction is
retained by the use of a magnetic separator. These are the CX3CR1+ 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.
Proper citation: Kelsey Miller 2016. MojoSort™ Mouse CX3CR1 Selection Kit Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.fyqbpvw Copy
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
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
Authors: Kelsey Miller
Group: BioLegend
Proper citation: Kelsey Miller 2017. ELISPOT Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.hv2b68e Copy
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
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
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
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
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
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
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
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
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
Authors: Kelsey Miller
Group: BioLegend
Proper citation: Kelsey Miller 2016. Nuclear Factor Fixation and Permeabilization Staining Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.e3bbgin Copy
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