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Mojosort mouse anti apc nanobeads

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Mojosort mouse anti-APC nanobeads are magnetic particles coated with antibodies specific to the APC (allophycocyanin) fluorescent protein. These nanobeads can be used to isolate and enrich cells expressing APC-labeled targets.

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Lab products found in correlation

Easysep mouse na ve cd8 t cell isolation kit, by STEMCELL (4 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions) Anti cd45 clone 30 f11, by BioLegend (1 mentions) Zombie aqua viability dye, by BioLegend (1 mentions) Anti epcam clone g8.8, by BioLegend (1 mentions) Anti epcam, by BioLegend (1 mentions) Triton x 100, by PerkinElmer (1 mentions) Glutaraldehyde, by Electron Microscopy Sciences (1 mentions) Siinfekl peptide, by Biosynth (1 mentions) Atp detection assay kit luminescence, by Cayman Chemical (1 mentions) Recombinant murine il 15, by Thermo Fisher Scientific (1 mentions) Poly d lysine, by Merck Group (1 mentions) Seahorse xf cell mito stress test kit, by Agilent Technologies (1 mentions) Facsaria iiiu, by BD (1 mentions) Ld column, by Miltenyi Biotec (1 mentions)

6 protocols using mojosort mouse anti apc nanobeads

1

Isolation of Lung Immune Cells

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Lung ILC2s (defined as Lin -CD45 + CD127 + CD90.2 + ST2 + CD25 + living cells) were sorted after negative enrichment against lineage markers (B220, CD11c, CD3, CD4, CD49b, CD5, CD8α, F4/80, FcεR1, Gr1 and Siglec-F) using MojoSort™ Mouse anti-APC Nanobeads (Biolegend) and magnetic separation using LD columns (Miltenyi). Lung ILCs (defined as Lin -CD45 + CD90.2 + living cells) were sorted after negative enrichment against lineage markers (B220, CD11c, CD3, CD4, CD5, CD8α, F4/80, FcεR1, Ly6G and Siglec-F) using MojoSort™ Mouse anti-APC Nanobeads and magnetic separation using LD columns (Miltenyi). AMs from BALF were defined as autofluorescent, FSC-A high living cells. Lung ECs were sorted (CD45-CD31-Epcam+ live cells) after depletion of CD45+ cells using using MojoSort™ Mouse anti-CD45 Nanobeads (Biolegend). BM Mos were sorted (CD19-CD3-CD11b+Ly6C+ living cells), after depletion of Ly6G+ and B220+ cells using MojoSort™ Mouse anti-APC Nanobeads (Biolegend) and magnetic separation using LD columns (Miltenyi). All cells were sorted on a FACSAria IIIu (BD Biosciences).
Loos P., Baiwir J., Maquet C., Javaux J., Sandor R., Lallemand F., Marichal T., Machiels B, & Gillet L. (2023). Dampening type 2 properties of group 2 innate lymphoid cells by a gammaherpesvirus infection reprograms alveolar macrophages. Science immunology, 8(80).
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2

Adoptive Transfer of Memory OT-I Cells

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Naïve CD8 T cells were harvested from CD45.1 OT-I mice (WT) or GZB-cre Zbtb20-fl/fl CD45.1 OT-I mice (KO) using EasySep mouse naïve CD8 T cell isolation kit (Stemcell Technologies cat:19858A). 50,000 naïve OT-I cells were retro-orbitally injected into B6 recipients, which were then retro-orbitally infected with 106 CFU LM-actA-Ova 1 day later. On D80 post infection, splenocytes were harvested from recipients, stained with anti-CD45.1-APC antibody then purified with Mojosort mouse anti-APC nanobeads (Biolegend Cat:480072). 106 enriched memory OT-I cells were adoptively transferred into MC38-Ova tumor-bearing mice, which were subcutaneously inoculated with 106 MC38-Ova tumor cells 4 days earlier. Tumor areas were measured three times a week.
Sun Y., Preiss N.K., Valenteros K.B., Kamal Y., Usherwood Y.K., Frost H.R, & Usherwood E.J. (2020). Zbtb20 restrains CD8 T cell immunometabolism and restricts memory differentiation and anti-tumor immunity. Journal of immunology (Baltimore, Md. : 1950), 205(10), 2649-2666.
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3

Flow Cytometry Analysis of Intestinal Epithelial Cells and Rotavirus Infection

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Dissociated cells were collected and stained for flow cytometry. Cells were stained with Zombie Aqua viability dye (BioLegend), Fc receptor-blocking antibody (CD16/CD32; BioLegend), anti-EpCAM (clone G8.8; BioLegend), and anti-CD45 (clone 30-F11; BioLegend). For analysis of murine rotavirus infection, cells were stained with anti-rotavirus (polyclonal; ThermoFisher, #PA1-7241) followed by goat anti-rabbit secondary (ThermoFisher). All data were analyzed using FlowJo software (BD Biosciences). Gates were set based on unstained and single-fluorophore stains. IECs were selected by gating on live, EpCAM-positive, CD45-negative cells. Gates for murine rotavirus infection were set based on naïve samples.
Where indicated, dissociated cells were enriched using MojoSort Mouse anti-APC Nanobeads (BioLegend, #480072) after flow cytometry staining for anti-EpCAM and anti-CD45 with APC fluorophores by following manufacturer protocols.
Van Winkle J.A., Peterson S.T., Kennedy E.A., Wheadon M.J., Ingle H., Desai C., Rodgers R., Constant D.A., Wright A.P., Li L., Artyomov M.N., Lee S., Baldridge M.T, & Nice T.J. (2022). Homeostatic interferon-lambda response to bacterial microbiota stimulates preemptive antiviral defense within discrete pockets of intestinal epithelium. eLife, 11, e74072.
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4

Measuring Metabolic Activity in Activated CD8 T Cells

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Naïve CD8 T cells were purified from spleens of CD45.1 OT-I mice (WT) or GZB-cre Zbtb20-fl/fl CD45.1 OT-I mice (KO) using EasySep mouse naïve CD8 T cell isolation kits (Stemcell Technologies cat:19858A). 50,000 naïve OT-I cells were retro-orbitally injected into congenic recipient mice, which were then retro-orbitally infected with 106 CFU LM-actA-Ova 1 day later. On D7 and D28 post infection, splenocytes were harvested from recipients, stained with anti-CD45.1-APC then purified with Mojosort mouse anti-APC nanobeads (Biolegend Cat:480072). Purified cells (purity greater than 95%) were then analyzed using a luminescence-based ATP detection assay (Cayman Chemical cat:700410).
Sun Y., Preiss N.K., Valenteros K.B., Kamal Y., Usherwood Y.K., Frost H.R, & Usherwood E.J. (2020). Zbtb20 restrains CD8 T cell immunometabolism and restricts memory differentiation and anti-tumor immunity. Journal of immunology (Baltimore, Md. : 1950), 205(10), 2649-2666.
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5

Metabolic Profiling of Antigen-Specific CD8+ T Cells

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Naïve CD8 T cells were harvested from CD45.1 OT-I mice (WT) or GZB-cre Zbtb20-fl/fl CD45.1 OT-I mice (KO) using EasySep mouse naïve CD8 T cell isolation kits (Stemcell Technologies cat:19858A). 50,000 naïve OT-I cells were retro-orbitally injected into B6 recipients, which were then retro-orbitally infected with 106 CFU LM-actA-Ova 1 day later. On D7 and D28 post infection, splenocytes were harvested from recipients, stained with anti-CD45.1-APC antibody then purified with Mojosort mouse anti-APC nanobeads (Biolegend Cat:480072). 200,000 enriched cells (purity greater than 95%) were seeded into each well for Seahorse mitochondrial stress tests and Glycolytic Rate tests. 1 μM oligomycin, 1.5 μM 4-(trifluoromethoxy)phenyl)carbonohydrazonoyl dicyanide (FCCP) and 0.5 μM Rotenone/Antimycin A were used for mitochondria stress assays. 0.5 μM Rotenone/Antimycin A and 50mM 2-deoxyglucose were used for Glycolytic rate assays.
Sun Y., Preiss N.K., Valenteros K.B., Kamal Y., Usherwood Y.K., Frost H.R, & Usherwood E.J. (2020). Zbtb20 restrains CD8 T cell immunometabolism and restricts memory differentiation and anti-tumor immunity. Journal of immunology (Baltimore, Md. : 1950), 205(10), 2649-2666.
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6

Murine CD8+ T Cell Isolation and Metabolic Profiling

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EasySep Mouse naïve CD8 T cell isolation kits (Stemcell Technologies cat:19858A); Mojosort mouse anti-APC nanobeads (Biolegend Cat:480072); ATP detection assay kit-luminescence (Cayman Chemical cat:700410); DAPI (Thermo Fisher cat:D1306); Seahorse XF Cell Mito Stress Test Kit (Seahorse Agilent cat:103015-100); 2-DG (Cayman Chemical cat:14325); SIINFEKL peptide (New England peptide Lot:V1355-37/40); recombinant human IL-2 (TECIN cat:Ro23-6019); recombinant murine IL-15 (PeproTech cat:210-15); poly-D-lysine (Millipore Sigma cat:P6407); Glutaraldehyde (Electron Microscopy Science cat:16000); NaBH4 (Alfa Aesar stock#:35788); Triton X-100 (PerkinElmer cat:N9300260).
Sun Y., Preiss N.K., Valenteros K.B., Kamal Y., Usherwood Y.K., Frost H.R, & Usherwood E.J. (2020). Zbtb20 restrains CD8 T cell immunometabolism and restricts memory differentiation and anti-tumor immunity. Journal of immunology (Baltimore, Md. : 1950), 205(10), 2649-2666.
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