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| Name | Authors | DOI | Group |
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MojoSort™ Human Pan Monocyte Isolation Kit Column Protocol Resource Report Resource Website |
Sam Li | DOI:10.17504/protocols.io.7tthnnn | 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 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. | BioLegend | https://www.biolegend.com/protocols/mojosort-human-pan-monocyte-isolation-kit-column-protocol/4765/ | 1 | 2019 | Sam Li 2019. MojoSort™ Human Pan Monocyte Isolation Kit Column Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.7tthnnn | 2021-04-15 09:15:16 | ||
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MojoSort™ Human anti-PE Nanobeads Protocol Resource Report Resource Website |
Sam Li | DOI:10.17504/protocols.io.7x3hpqn | BioLegend | Product description and procedure summary: Target cells are positively selected or depleted by incubating the sample with an anti-human PE conjugated antibody, followed by incubation with magnetic anti-PE Nanobeads. The magnetically labeled fraction is retained by the use of a magnetic separator. These are the PE+ 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.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. | BioLegend | https://www.biolegend.com/protocols/mojosort-human-anti-pe-nanobeads-protocol/4753/ | 1 | 2019 | Sam Li 2019. MojoSort™ Human anti-PE Nanobeads Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.7x3hpqn | 2021-04-15 09:15:49 | ||
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Intracellular Flow Cytometry Staining Protocol Resource Report Resource Website |
Sam Li | DOI:10.17504/protocols.io.bac7iazn | BioLegend | BioLegend | https://www.biolegend.com/protocols/intracellular-flow-cytometry-staining-protocol/4260/ | 3 | 2019 | Sam Li 2019. Intracellular Flow Cytometry Staining Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.bac7iazn | 2021-04-15 09:15:29 | |||
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Chromatin Immunoprecipitation (ChIP) Assay Protocol Resource Report Resource Website |
Sam Li | DOI:10.17504/protocols.io.gsmbwc6 | BioLegend | Reagents - Basic cell culture media: respective to the cell line used and experimental design Distilled water: Invitrogen® UltraPure™ Distilled Water (catalog number 19977015) or equivalent product from other vendor.Highthroughput (HT) Pro A or G 96 well plate or Spin Column (Chromatrap® HT ChIP-qPCR kit; catalog number 500161 or 500162 or 500163)96-well collection plate: USA scientific® TempPlate non-skirted 96-well PCR plate natural (catalog number 1402-9596) or equivalent product from other vendor.QIAquick™ PCR purification kit (Qiagen®; catalog number 28104) or equivalent product from other vendor.Hypotonic Buffer (Chromatrap® HT ChIP-qPCR kit; catalog number 500161 or 500162 or 500163)Digestion Buffer (Chromatrap® HT ChIP-qPCR kit; catalog number 500161 or 500162 or 500163)Lysis Buffer (Chromatrap® HT ChIP-qPCR kit; catalog number 500161 or 500162 or 500163)Column Conditioning Buffer (Chromatrap® HT ChIP-qPCR kit; catalog number 500161 or 500162 or 500163)Wash Buffer 1 (Chromatrap® HT ChIP-qPCR kit; catalog number 500161 or 500162 or 500163)Wash Buffer 2 (Chromatrap® HT ChIP-qPCR kit; catalog number 500161 or 500162 or 500163)Wash Buffer 3 (Chromatrap® HT ChIP-qPCR kit; catalog number 500161 or 500162 or 500163)Elution Buffer (Chromatrap® HT ChIP-qPCR kit; catalog number 500161 or 500162 or 500163)5M NaCl: dissolve 292g of NaCl in 800mL of H2O. Adjust the volume to 1L with H2O.1M NaHCO3: dissolve 12.6g of NaHCO3 in 100 mL of H2O. Adjust the volume to 150 mL with H2O.Protease Inhibitor Cocktail (PIC): Cell signaling Technology® (catalog number 5871) or equivalent product from other vendor.Proteinase K: Qiagen® (catalog number 19133) or equivalent product from other vendor.Proteinase K stop solution: 100mM Phenylmethylsulfonyl fluoride (PMSF). Weigh 1.742g PMSF (Sigma-Aldrich®, catalog number 10837091001) and add DMSO (Sigma-Aldrich®, catalog number D2650) to a final volume of 100mL. Dissolve completely.PBS: HyClone™ Phosphate Buffered Saline (1X) (catalog number SH30256.01) or equivalent product from other vendor.Formaldehyde: VWR® 37% w/w aqueous solution (catalog number 97064-604) or equivalent product from other vendor.0.65M Glycine: Dissolve 488g of glycine into 1L of H2O to make 6.5M glycine (10X). Dilute with PBS to make 1X before each experiment. Micrococcal Nuclease: Cell Signaling Technologies® (catalog number 10011S) or equivalent product from other vendor.1.3-1.5% Agarose gel: dissolve 1.3-1.5g of agarose powder into 100mL of TAE buffer by microwave heating. Visualize DNA under UV light by adding appropriate amount of Ethidium Bromide, or use an equivalent method.Add appropriate amount of Ethidium Bromide to cooled down gel solution to visualize DNA under UV light.0.5M EDTA: Dissolve 186.1g EDTA into 700mL of H2O. Adjust the volume to 1L with H2O.100bp ladder: NEB® (catalog number N0467S) or equivalent product from other vendor.Materials -Heat blockRotatorCentrifugeMicrocentrifuge37ºC incubatorNanodropSonicatorUV light imagerRocking platform | BioLegend | http://www.biolegend.com/media_assets/support_protocol/ChIP_Assay_Technical_Protocol_v8.pdf | 1 | 2016 | Sam Li 2016. Chromatin Immunoprecipitation (ChIP) Assay Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.gsmbwc6 | 2021-04-15 09:15:33 | ||
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LEGENDplex™ Assay Setup Protocol for Cytek® Aurora and Northern Lights Flow Cytometers Resource Report Resource Website |
Sam Li | DOI:10.17504/protocols.io.bi3mkgk6 | BioLegend | To view images that accompany this protocol, click here.This protocol provides step-by-step instructions for the setup of Cytek® Aurora and Northern Lights flow cytometers for data acquisition of BioLegend’s LEGENDplex™ multiplex immunoassays. A pre-made LEGENDplex™ Experiment Template that can easily be imported into the SpectroFlo® software is available for download at biolegend.com/en-us/legendplex under the Instrument Setup tab. If desired, users can also create their own Experiment Template by following the instructions found at the end of this protocol. Unmixing of spectral data is not required. Please export Raw FCS files for downstream analysis. This is analogous to using uncompensated data on a conventional flow cytometer. When analyzing LEGENDplex™ Assay FCS files generated on an Aurora or Northern Lights flow cytometer, the assay beads emission spectra should be analyzed using the following recommended channels: • Without a yellow-green (YG) laser: Bead Classification = R4, Reporter = B4 • With a yellow-green (YG) laser: Bead Classification = R4, Reporter = YG1 | BioLegend | https://www.biolegend.com/Files/Images/BioLegend/legendplex/instructions/Setup_Procedure_for_Cytek_Aurora_and_Northern_Lights_2_Lasers.pdf | 1 | 2020 | Sam Li 2020. LEGENDplex™ Assay Setup Protocol for Cytek® Aurora and Northern Lights Flow Cytometers. protocols.io https://dx.doi.org/10.17504/protocols.io.bi3mkgk6 | 2021-04-15 09:15:52 | ||
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Th17 Polarization of Mouse CD4+ Cells Resource Report Resource Website |
Sam Li | DOI:10.17504/protocols.io.79vhr66 | BioLegend | BioLegend | https://www.biolegend.com/protocols/th17-polarization-of-mouse-cd4--cells/4284/ | 3 | 2019 | Sam Li 2019. Th17 Polarization of Mouse CD4+ Cells. protocols.io https://dx.doi.org/10.17504/protocols.io.79vhr66 | 2021-04-15 09:15:52 | |||
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MojoSort™ Human CD41 Nanobeads Protocol Resource Report Resource Website |
Sam Li | DOI:10.17504/protocols.io.7xxhppn | BioLegend | Product description and procedure summary: 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 14 mL tubes and Magnet, and place the tube in the magnet for 10 minutes.Application notes: To use this product in magnetic separation columns, a titration of the Nanobeads should be performed. Optimal concentration for magnetic separation columns is lot-specific. Please contact BioLegend Technical Service ([email protected]) for further assistance on how to use MojoSort™ Nanobeads in magnetic separation columns. | BioLegend | https://www.biolegend.com/protocols/mojosort-human-cd41-nanobeads-protocol/4535/ | 1 | 2019 | Sam Li 2019. MojoSort™ Human CD41 Nanobeads Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.7xxhppn | 2021-04-15 09:15:53 | ||
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MojoSort™ Human CD4 T Cell Selection Kit Column Protocol - CD14 Selection Resource Report Resource Website |
Sam Li | DOI:10.17504/protocols.io.7txhnpn | 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 than 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. | BioLegend | https://www.biolegend.com/protocols/mojosort-human-cd4-t-cell-selection-kit-column-protocol-cd14-selection/4763/ | 1 | 2019 | Sam Li 2019. MojoSort™ Human CD4 T Cell Selection Kit Column Protocol - CD14 Selection. protocols.io https://dx.doi.org/10.17504/protocols.io.7txhnpn | 2021-04-15 09:15:52 | ||
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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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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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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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Flex-T™ Fixed Peptide Tetramer Preparation and Flow Cytometry Staining Protocol Resource Report Resource Website |
Sam Li | BioLegend | BioLegend | https://www.biolegend.com/en-us/protocols/flex-t-fixed-peptide-tetramer-preparation-and-flow-cytometry-staining-protocol | 1 | 2020 | Sam Li 2020. Flex-T™ Fixed Peptide Tetramer Preparation and Flow Cytometry Staining Protocol. protocols.io https:// | 2021-04-15 09:16:27 | ||||
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MojoSort™ Mouse anti-APC Nanobeads Column Protocol Resource Report Resource Website |
Sam Li | DOI:10.17504/protocols.io.699hh96 | 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 than 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. | BioLegend | https://www.biolegend.com/protocols/mojosort-mouse-anti-apc-nanobeads-column-protocol/4759/ | 1 | 2019 | Sam Li 2019. MojoSort™ Mouse anti-APC Nanobeads Column Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.699hh96 | 2021-04-15 09:16:16 | ||
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MojoSort™ Human NK Cell Isolation Kit Protocol Resource Report Resource Website |
Sam Li | DOI:10.17504/protocols.io.7bihike | 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 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. | BioLegend | https://www.biolegend.com/protocols/mojosort-human-nk-cell-isolation-kit-column-protocol/4622/ | 1 | 2019 | Sam Li 2019. MojoSort™ Human NK Cell Isolation Kit Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.7bihike | 2021-04-15 09:16:33 | ||
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Immunocytochemistry Staining Protocol Resource Report Resource Website |
Sam Li | DOI:10.17504/protocols.io.968h9hw | BioLegend | BioLegend | https://www.biolegend.com/protocols/immunocytochemistry-staining-protocol/4275/ | 4 | 2019 | Sam Li 2019. Immunocytochemistry Staining Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.968h9hw | 2021-04-15 09:16:21 | |||
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MojoSort™ Isolation Kits Column Protocol - 3 Resource Report Resource Website |
Sam Li | DOI:10.17504/protocols.io.7behije | 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 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. | BioLegend | https://www.biolegend.com/protocols/mojosort-isolation-kits-column-protocol-3/4591/ | 1 | 2019 | Sam Li 2019. MojoSort™ Isolation Kits Column Protocol - 3. protocols.io https://dx.doi.org/10.17504/protocols.io.7behije | 2021-04-15 09:16:40 | ||
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MojoSort™ Streptavidin Nanobeads Protocol - Positive Selection Resource Report Resource Website |
Sam Li | DOI:10.17504/protocols.io.7wghpbw | BioLegend | Product description and procedure summary:If your target cells are the labeled cells (the positive fraction), use the Streptavidin Nanobeads Protocol – Positive Selection. If your target cells are the unlabeled cells (negative fraction), use the Streptavidin Nanobeads Protocol - Negative Selection. Target cells are isolated by incubating your sample with a Biotin-conjugated antibody or 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. | BioLegend | https://www.biolegend.com/protocols/mojosort-streptavidin-nanobeads-protocol-positive-selection/4748/ | 1 | 2019 | Sam Li 2019. MojoSort™ Streptavidin Nanobeads Protocol - Positive Selection. protocols.io https://dx.doi.org/10.17504/protocols.io.7wghpbw | 2021-04-15 09:16:25 | ||
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Chromatin Immunoprecipitation (ChIP) Assay Protocol Resource Report Resource Website |
Sam Li | DOI:10.17504/protocols.io.97gh9jw | BioLegend | BioLegend | https://www.biolegend.com/protocols/chromatin-immunoprecipitation-chip-assay-protocol/4306/ | 4 | 2019 | Sam Li 2019. Chromatin Immunoprecipitation (ChIP) Assay Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.97gh9jw | 2021-04-15 09:16:45 | |||
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MojoSort™ Mouse CD326 (Ep-CAM) Selection Kits Column Protocol Resource Report Resource Website |
Sam Li | DOI:10.17504/protocols.io.7u4hnyw | 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 and less antibody than 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. | BioLegend | https://www.biolegend.com/en-us/protocols/mojosort-mouse-cd326-ep-cam-selection-kits-column-protocol | 1 | 2019 | Sam Li 2019. MojoSort™ Mouse CD326 (Ep-CAM) Selection Kits Column Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.7u4hnyw | 2021-04-15 09:16:43 |
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