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| Name | Authors | DOI | Group |
Summary |
Associated Publications |
RRIDs used | ||||||
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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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Polarization of Mouse CD4+ Cells Resource Report Resource Website |
BioLegend, Inc. | DOI:10.17504/protocols.io.ezsbf6e | BioLegend | This is collection of BioLegend protocols for polarization of mouse CD4+ cells. Please make sure you refer to the correct protocol, depending on whether you are using Th1, Th2, Th17, or Treg. | , | 1 | 2016 | BioLegend, Inc. 2016. Polarization of Mouse CD4+ Cells. protocols.io https://dx.doi.org/10.17504/protocols.io.ezsbf6e | 2021-04-15 09:15:57 | |||
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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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Anti-c-Myc Tag (9E10) Affinity Gel Protocol Resource Report Resource Website |
Kelsey Knight | DOI:10.17504/protocols.io.tideka6 | BioLegend | Catalog Number: 658502 Storage Temperature: 2°C-8°C Product Description Anti-c-Myc Tag (9E10) Affinity Gel consists of anti-c-myc monoclonal antibody (clone 9E10), covalently immobilized onto 6% high density glyoxal agarose beads. The affinity resin can be used in affinity purifying and immunoprecipitation of c-Myc-tagged fusion protein. Binding Specificity Mouse monoclonal antibody 9E10 recognizes the c-myc epitope N-EQKLSEEDL-C and is purified through Protein G chromatography. The antibody conjugated affinity resin can bind to epitope at N-terminal, C-terminal, and internal locations of a fusion protein. The binding capacity is greater than 0.5mg per ml of c-myc agarose resin. Reagent Anti-c-Myc Agarose Affinity Gel is supplied as a 50% suspension in phosphate-buffered saline, pH 7.2, containing 0.09% sodium azide. Precautions and Disclaimer This product is for research use only. Please consult the Material Safety Data Sheet for information regarding hazards and safe handling practices. Storage/Stability Anti-c-Myc Agarose Affinity Gel should be stored between 2°C and 8°C for maximum stability. The unopened product is stable for one year when stored as indicated. After use, the resin should be regenerated and stored in TBS or PBS buffer (pH 7.2) containing 0.09% sodium azide. | BioLegend | https://www.biolegend.com/protocols/anti-c-myc-tag-9e10-affinity-gel-protocol/4297/ | 1 | 2018 | Kelsey Knight 2018. Anti-c-Myc Tag (9E10) Affinity Gel Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.tideka6 | 2021-04-15 09:15:59 | ||
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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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Immunohistochemistry Protocol for Frozen Sections Resource Report Resource Website |
BioLegend, Inc. | DOI:10.17504/protocols.io.eywbfxe | BioLegend | The following is a general procedure guide for preparation and staining of acetone-fixed frozen tissues using a purified, unconjugated primary antibody, biotinylated secondary antibody and streptavidin-horseradish peroxidase (Sav-HRP) and DAB detection system. Because each antigen differs in terms of requirement for fixation, amplification step, etc., it is not possible to write an inclusive protocol that will work for all antigens. The user must determine optimal conditions for each antigen of interest. Many protocols for staining individual antigens, as well as useful tips and troubleshooting guides for immunohistochemistry, can be found at the IHC World web site (http://www.ihcworld.com/). | , | http://www.biolegend.com/media_assets/support_protocol/BioLegend_IHC_Frozen_121511.pdf | 1 | 2016 | BioLegend, Inc. 2016. Immunohistochemistry Protocol for Frozen Sections. protocols.io https://dx.doi.org/10.17504/protocols.io.eywbfxe | 2021-04-15 09:16:05 | ||
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Anti-BrdU Staining Using 70% Ethanol and 2N HCl Resource Report Resource Website |
Sam Li | DOI:10.17504/protocols.io.bac8iazw | BioLegend | Note: We offer two protocols here depending on what your experiment requires. Ethanol treatment is usually harsher toward any other fluors or fluorescent proteins that may be present in your sample. As such, the DNAse method may be gentler under those conditions | BioLegend | https://www.biolegend.com/protocols/anti-brdu-staining-using-70-ethanol-and-2n-hcl/4239/ | 4 | 2019 | Sam Li 2019. Anti-BrdU Staining Using 70% Ethanol and 2N HCl. protocols.io https://dx.doi.org/10.17504/protocols.io.bac8iazw | 2021-04-15 09:16:05 | ||
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True-Nuclear™ Transcription Factor Staining Protocol for 5 mL Tubes Resource Report Resource Website |
Kelsey Knight | DOI:10.17504/protocols.io.tqxemxn | BioLegend | BioLegend | https://www.biolegend.com/protocols/true-nuclear-transcription-factor-staining-protocol-for-5ml-tubes/4267/ | 3 | 2018 | Kelsey Knight 2018. True-Nuclear™ Transcription Factor Staining Protocol for 5 mL Tubes. protocols.io https://dx.doi.org/10.17504/protocols.io.tqxemxn | 2021-04-15 09:16:09 | |||
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Anti-Neu5Gc Antibody Kit Protocol - Western Blot Resource Report Resource Website |
Sam Li | DOI:10.17504/protocols.io.97hh9j6 | BioLegend | The Anti-Neu5Gc Antibody Kit contains the essential monospecific polyclonal chicken IgY antibody, along with a negative control primary antibody to detect the presence of Neu5Gc on glycoconjugates by Western blot (WB). Samples to be evaluated are first subjected to SDS-PAGE, followed by transfer to a nitrocellulose or polyvinylidenedifluoride (PVDF) membrane. The membrane is then incubated with affinity-purified polyclonal anti-Neu5Gc to determine the presence of Neu5Gc on the protein of interest.The antibody provided in this kit has been shown to identify as little as 5pmol of Neu5Gc per µg glycoprotein, which is at or below the current detection limit for conventional analysis by acid release, purification, DMB derivatization, HPLC, and electrospray mass-spectrometry. The Western blot provides additional information in that it confirms that Neu5Gc is directly linked to the glycoprotein of interest rather than to an accompanying sample component. | BioLegend | https://www.biolegend.com/protocols/anti-neu5gc-antibody-kit-protocol-western-blot/4300/ | 3 | 2019 | Sam Li 2019. Anti-Neu5Gc Antibody Kit Protocol - Western Blot. protocols.io https://dx.doi.org/10.17504/protocols.io.97hh9j6 | 2021-04-15 09:16:12 | ||
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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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Immunoprecipitation Protocol Resource Report Resource Website |
Sam Li | DOI:10.17504/protocols.io.98qh9vw | BioLegend | Immunoprecipitation is a procedure by which proteins or peptides that react specifically with an antibody are removed from solution and examined for quantity or physical characteristics. Immunoprecipitation can also be used to “enrich” a protein population prior to Western Blotting. For example, one can perform immunoprecipitation with a pan-specific antibody against a protein of interest followed by Western blotting with a modification-specific antibody (such as a phospho-specific antibody or an acetylation-specific antibody). | BioLegend | https://www.biolegend.com/protocols/immunoprecipitation-protocol/4258/ | 3 | 2019 | Sam Li 2019. Immunoprecipitation Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.98qh9vw | 2021-04-15 09:16:23 | ||
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MojoSort™ Selection Kits Protocol - 1 Resource Report Resource Website |
Sam Li | DOI:10.17504/protocols.io.7yxhpxn | BioLegend | 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 14mL tubes and Magnet, and place the tube in the magnet for 10 minutes. | BioLegend | https://www.biolegend.com/protocols/mojosort-selection-kits-protocol-1/4657/ | 2 | 2019 | Sam Li 2019. MojoSort™ Selection Kits Protocol - 1. protocols.io https://dx.doi.org/10.17504/protocols.io.7yxhpxn | 2021-04-15 09:16:28 | ||
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Cell Surface Immunofluorescence Staining Protocol Resource Report Resource Website |
BioLegend, Inc. | DOI:10.17504/protocols.io.hvyb67w | BioLegend | , | http://www.biolegend.com/media_assets/support_protocol/BioLegend_Surface_Staining_Flow_Protocol_060215.pdf | 2 | 2017 | BioLegend, Inc. 2017. Cell Surface Immunofluorescence Staining Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.hvyb67w | 2021-04-15 09:16:26 |
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