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| Name | Authors | DOI | Group |
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Associated Publications |
RRIDs used | ||||||
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Flex-T™ HLA Class I ELISA Protocol Resource Report Resource Website |
Kelsey Miller | DOI:10.17504/protocols.io.hv4b68w | BioLegend | 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. | BioLegend | http://www.biolegend.com/media_assets/support_protocol/Protocol%20for%20HLA%20class%20I%20ELISA%2006202016.pdf | 2 | 2017 | Kelsey Miller 2017. Flex-T™ HLA Class I ELISA Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.hv4b68w | 2021-04-15 09:15:27 | ||
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Immunohistochemistry Protocol for Ultra Streptavidin Detection Kits (USA) Resource Report Resource Website |
Kelsey Miller | DOI:10.17504/protocols.io.fq9bmz6 | BioLegend | BioLegend | http://www.biolegend.com/media_assets/support_protocol/IHC_Protocol_for_Ultra_Streptavidin_Detection_Kits_USA_090815.pdf | 2 | 2016 | Kelsey Miller 2016. Immunohistochemistry Protocol for Ultra Streptavidin Detection Kits (USA). protocols.io https://dx.doi.org/10.17504/protocols.io.fq9bmz6 | 2021-04-15 09:15:25 | |||
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Immunohistochemistry Protocol for Beta Amyloid Products using USA Detection Kit Resource Report Resource Website |
Kelsey Miller | DOI:10.17504/protocols.io.e2ibgce | BioLegend | BioLegend | http://www.biolegend.com/media_assets/support_protocol/Beta_Amyloid_IHC_Protocol_using_USA_Kit_10-29-14.pdf | 1 | 2016 | Kelsey Miller 2016. Immunohistochemistry Protocol for Beta Amyloid Products using USA Detection Kit. protocols.io https://dx.doi.org/10.17504/protocols.io.e2ibgce | 2021-04-15 09:15:48 | |||
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Immunofluorescence Microscopy Protocol with Methanol Fixed Cells Resource Report Resource Website |
Kelsey Miller | DOI:10.17504/protocols.io.hv9b696 | BioLegend | BioLegend | 2 | 2017 | Kelsey Miller 2017. Immunofluorescence Microscopy Protocol with Methanol Fixed Cells. protocols.io https://dx.doi.org/10.17504/protocols.io.hv9b696 | 2021-04-15 09:15:35 | ||||
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Flex-T™ HLA Class I ELISA Protocol Resource Report Resource Website |
Kelsey Miller | DOI:10.17504/protocols.io.e7ebhje | BioLegend | 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. | BioLegend | http://www.biolegend.com/media_assets/support_protocol/Protocol%20for%20HLA%20class%20I%20ELISA%2006202016.pdf | 1 | 2016 | Kelsey Miller 2016. Flex-T™ HLA Class I ELISA Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.e7ebhje | 2021-04-15 09:15:42 | ||
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Immunohistochemistry Protocol for Keratin Antibodies Resource Report Resource Website |
Kelsey Miller | DOI:10.17504/protocols.io.tkhekt6 | BioLegend | BioLegend | https://www.biolegend.com/protocols/immunohistochemistry-protocol-for-keratin-antibodies/4287/ | 2 | 2018 | Kelsey Miller 2018. Immunohistochemistry Protocol for Keratin Antibodies. protocols.io https://dx.doi.org/10.17504/protocols.io.tkhekt6 | 2021-04-15 09:15:40 | |||
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Cell Fixation and Permeabilization Protocol using 70% Ethanol Resource Report Resource Website |
Kelsey Miller | DOI:10.17504/protocols.io.hv3b68n | BioLegend | Cell Fixation and Permeabilization Protocol Using 70% Ethanol | BioLegend | http://www.biolegend.com/media_assets/support_protocol/BioLegend_EthFIXPERM_032912.pdf | 2 | 2017 | Kelsey Miller 2017. Cell Fixation and Permeabilization Protocol using 70% Ethanol. protocols.io https://dx.doi.org/10.17504/protocols.io.hv3b68n | 2021-04-15 09:15:53 | ||
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ACUITYAdvanced Protocol Resource Report Resource Website |
Kelsey Miller | DOI:10.17504/protocols.io.e37bgrn | BioLegend | ACUITYAdvanced Biotin Free HRP Polymer Detection System for Immunohistochemistry | BioLegend | http://www.biolegend.com/media_assets/support_protocol/ACUITYAdvanced_Protocol_052115.pdf | 1 | 2016 | Kelsey Miller 2016. ACUITYAdvanced Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.e37bgrn | 2021-04-15 09:15:56 | ||
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MojoSort™ Isolation Kits Regular Protocol Resource Report Resource Website |
Kelsey Miller | DOI:10.17504/protocols.io.e2vbge6 | BioLegend | 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 your cells of interest; do not discard the liquid. Some of the downstream applications include functional assays, gene expression, phenotypic characterization, etc. | BioLegend | http://www.biolegend.com/media_assets/support_protocol/MojoSort_Isolation_Kits_Regular_Protocol_V02_10192015.pdf | 1 | 2016 | Kelsey Miller 2016. MojoSort™ Isolation Kits Regular Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.e2vbge6 | 2021-04-15 09:16:00 | ||
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Th9 Polarization of Mouse CD4+ Cells Resource Report Resource Website |
Kelsey Miller | DOI:10.17504/protocols.io.tnnemde | BioLegend | BioLegend | https://www.biolegend.com/protocols/th9-polarization-of-mouse-cd4--cells-protocol/4243/ | 2 | 2018 | Kelsey Miller 2018. Th9 Polarization of Mouse CD4+ Cells. protocols.io https://dx.doi.org/10.17504/protocols.io.tnnemde | 2021-04-15 09:16:06 | |||
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Anti-BrdU Staining Protocols Using DNAse with Surface and Fluorescent Proteins Resource Report Resource Website |
Kelsey Miller | DOI:10.17504/protocols.io.hvkb64w | BioLegend | Anti-BrdU Staining Protocol using DNAse with surface and fluorescent proteins | BioLegend | http://www.biolegend.com/media_assets/support_protocol/BrdU_Staining_Protocols_05102016.pdf | 2 | 2017 | Kelsey Miller 2017. Anti-BrdU Staining Protocols Using DNAse with Surface and Fluorescent Proteins. protocols.io https://dx.doi.org/10.17504/protocols.io.hvkb64w | 2021-04-15 09:16:10 | ||
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Immunopurification — Small Scale using Epitope Tag Affinity Matrices Resource Report Resource Website |
Kelsey Miller | DOI:10.17504/protocols.io.e36bgre | BioLegend | Immunochemistry Protocols: Immunopurification ‐ (Small Scale) Using Anti‐Epitope Tag Affinity Matrices:Typically, immunoprecipitation of antigen from extracts of transfected mammalian cells requires between 1-5 μL of polyclonal antiserum, 5-100 μL of hybridoma tissue culture medium, or 1-3 μL of ascites. If more antibody is used than is necessary, nonspecific background will increase. The concentration of immunoglobulins in antiserum is approximately 6-8 mg/mL. Supernatant from cultured hybridoma cells typically contains 10-100 μg/mL immunoglobulin, and ascites fluid contains 5-7 mg/mL immunoglobulin. | BioLegend | http://www.biolegend.com/media_assets/support_protocol/Immunopurification_Sm_Scale_Epitope_Tag_Affinity_Matrices.pdf | 1 | 2016 | Kelsey Miller 2016. Immunopurification — Small Scale using Epitope Tag Affinity Matrices. protocols.io https://dx.doi.org/10.17504/protocols.io.e36bgre | 2021-04-15 09:16:10 | ||
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MojoSort™ Human CD4 Memory T Cells Isolation Kit Protocol Resource Report Resource Website |
Kelsey Miller | DOI:10.17504/protocols.io.fxqbpmw | BioLegend | 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.Product description and procedure summary: The kit is designed for the isolation of human memory CD4 T cells 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. | BioLegend | http://www.biolegend.com/media_assets/support_protocol/MojoSort_CD4_Memory_T_Cell_Protocol_08122016.pdf | 1 | 2016 | Kelsey Miller 2016. MojoSort™ Human CD4 Memory T Cells Isolation Kit Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.fxqbpmw | 2021-04-15 09:16:12 | ||
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Anti-Neu5Gc Antibody Kit Protocol - ELISA Resource Report Resource Website |
Kelsey Miller | DOI:10.17504/protocols.io.tigekbw | BioLegend | 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. | BioLegend | https://www.biolegend.com/protocols/anti-neu5gc-antibody-kit-protocol-elisa/4299/ | 3 | 2018 | Kelsey Miller 2018. Anti-Neu5Gc Antibody Kit Protocol - ELISA. protocols.io https://dx.doi.org/10.17504/protocols.io.tigekbw | 2021-04-15 09:16:11 | ||
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Anti-Neu5Gc Antibody Kit Protocol - Flow Cytometry Resource Report Resource Website |
Kelsey Miller | DOI:10.17504/protocols.io.e26bghe | BioLegend | Anti-Neu5Gc may be used for staining cells prior to analysis by Flow Cytometry. This kit contains all the essential components needed to identify Neu5Gc on the surface of cells by flow cytometry.Use of the blocking agent (Neu5Gc Assay Blocking Solution) provided in the kit is essential, as commonly used blocking agents invariably contain serum, or serum components, that can either inhibit detection or introduce Neu5Gc contamination.Tissue culture-grown CHO-K1 can be used as a positive control, and human peripheral blood mononuclear cells serve as a negative control. Adherent tissue culture-grown cells should be released from the culture flasks by using 5-10mM EDTA for 10 minutes at room temperature. Other non-enzymatic methods, such as Accutase, may be used. Immediately wash cells in blocking buffer that contains a lower concentration of EDTA, and resuspend cells in blocking buffer to determine cell numbers and viability. | BioLegend | http://www.biolegend.com/media_assets/support_protocol/Neu5Gc_Antibody_Kit_Protocols_110813.pdf | 1 | 2016 | Kelsey Miller 2016. Anti-Neu5Gc Antibody Kit Protocol - Flow Cytometry. protocols.io https://dx.doi.org/10.17504/protocols.io.e26bghe | 2021-04-15 09:16:17 | ||
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Immunofluorescent Staining of Foxp3 in Frozen Sections Resource Report Resource Website |
Kelsey Miller | DOI:10.17504/protocols.io.e3pbgmn | BioLegend | T regulatory cells (also known as Tregs or Regulatory T cells) are essential cells in the immune system that suppress immune responses of other cells, designed to limit excessive reactions and prevent autoimmunity. Tregs are characterized by the expression of CD4, CD25, and Foxp3, while lacking CD127. CD4+Foxp3+ regulatory T cells have been referred to as “naturally-occurring” regulatory T cells to distinguish them from “suppressor” T cell populations that are generated in vitro. While other variants of suppressive T cells do exist, such as CD8 suppressor cells, Th3 and Tr-1 cells, Tregs are classically defined as CD4+CD25+Foxp3+ cells. Here, we provide a protocol for immunofluorescent staining of FOXP3 in frozen sections, adapted from a protocol from Dr. Matthias Hardtke-Wolenski, Medizinische Hochschule Hannover. This protocol has been successfully used on mouse spleen, liver, and skin sections. | BioLegend | http://www.biolegend.com/media_assets/support_protocol/BioLegend_IF-FOXP3_121511.pdf | 1 | 2016 | Kelsey Miller 2016. Immunofluorescent Staining of Foxp3 in Frozen Sections. protocols.io https://dx.doi.org/10.17504/protocols.io.e3pbgmn | 2021-04-15 09:16:16 | ||
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Anti-Neu5Gc Antibody Kit Protocol - Flow Cytometry Resource Report Resource Website |
Kelsey Miller | DOI:10.17504/protocols.io.hvqb65w | BioLegend | Anti-Neu5Gc may be used for staining cells prior to analysis by Flow Cytometry. This kit contains all the essential components needed to identify Neu5Gc on the surface of cells by flow cytometry.Use of the blocking agent (Neu5Gc Assay Blocking Solution) provided in the kit is essential, as commonly used blocking agents invariably contain serum, or serum components, that can either inhibit detection or introduce Neu5Gc contamination.Tissue culture-grown CHO-K1 can be used as a positive control, and human peripheral blood mononuclear cells serve as a negative control. Adherent tissue culture-grown cells should be released from the culture flasks by using 5-10mM EDTA for 10 minutes at room temperature. Other non-enzymatic methods, such as Accutase, may be used. Immediately wash cells in blocking buffer that contains a lower concentration of EDTA, and resuspend cells in blocking buffer to determine cell numbers and viability. | BioLegend | http://www.biolegend.com/media_assets/support_protocol/Neu5Gc_Antibody_Kit_Protocols_110813.pdf | 2 | 2017 | Kelsey Miller 2017. Anti-Neu5Gc Antibody Kit Protocol - Flow Cytometry. protocols.io https://dx.doi.org/10.17504/protocols.io.hvqb65w | 2021-04-15 09:16:14 | ||
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MojoSort™ Positive Selection Columns Protocol Resource Report Resource Website |
Kelsey Miller | DOI:10.17504/protocols.io.e3kbgkw | BioLegend | 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. | BioLegend | 1 | 2016 | Kelsey Miller 2016. MojoSort™ Positive Selection Columns Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.e3kbgkw | 2021-04-15 09:16:20 | |||
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Propidium Iodide Cell Cycle Staining Protocol Resource Report Resource Website |
Kelsey Miller | DOI:10.17504/protocols.io.tm8ek9w | BioLegend | BioLegend | https://www.biolegend.com/protocols/propidium-iodide-cell-cycle-staining-protocol/4303/ | 2 | 2018 | Kelsey Miller 2018. Propidium Iodide Cell Cycle Staining Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.tm8ek9w | 2021-04-15 09:16:40 | |||
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MojoSort™ Human CD4 Nanobeads Regular Protocol Resource Report Resource Website |
Kelsey Miller | DOI:10.17504/protocols.io.e3hbgj6 | BioLegend | This procedure is optimized for the isolation of 107 to 2 x 108 cells per tube from human peripheral blood mononuclear cells (PBMCs). 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. To maximize the purity of CD4+ T cells we recommend the depletion of monocytes, as human monocytes express CD4. To deplete monocytes from human PBMCs, we recommend the use of MojoSort™ Human CD14 Selection Kit (Cat. No. 480025/480026). Alternatively, you may consider the Human CD4 T Cell Selection Kit (Cat. No. 480038). After monocyte depletion proceed to isolate the CD4+ T cells. You can also use other methods to deplete monocytes. If you wish to include monocytes in your CD4+ cell isolation, depletion is not required. | BioLegend | http://www.biolegend.com/media_assets/support_protocol/MojoSort_Human_CD4_Nanobeads_Protocol_V02_10192015.pdf | 1 | 2016 | Kelsey Miller 2016. MojoSort™ Human CD4 Nanobeads Regular Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.e3hbgj6 | 2021-04-15 09:16:29 |
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