Mice were injected intraperitoneally with 10 mg/kg of LPS from E. coli O111:B4 (Sigma-Aldrich, St. Louis, MO) or 7.5 μg of recombinant TNF (Peprotech, East Windsor, NJ) and euthanized after 90 min. Where indicated, mice were pre-treated with 150 μg anti-CD103 (M290, BioXcell, Lebanon, NH) or 200 μg of anti-NKG2D (HMG2D, BioXcell) 1 h prior to LPS treatment; or 2 × 200 μg anti-TCRγδ (UC7-13D5, BioXcell) 24 and 48h prior to LPS treatment. Rat IgG2a (RTK2758, BioLegend) or Armenian hamster IgG (BioXcell) were used as controls. 100 μg propidium iodide (Thermo Fisher Scientific, Beverly, MA) was administered i.v. prior to euthanasia. As a control for pyroptosis, 0.16 μg/g PA and 0.08 μg/g LFn-Fla (L. pneumophila flagellin, provided by Isabella Rauch (OHSU)) was administered i.v. for 60 min20 (link). To quantify shedding events, immunohistochemistry was performed as described in supplementary materials . Positive shedding events were identified via ZO-1 funnels, CC3 staining and/or cellular morphology.
Rat igg2a rtk2758
Manufactured by BioLegend
Rat IgG2a (RTK2758) is an antibody isotype control reagent. It is designed to be used as a non-specific negative control in flow cytometry, cell-based assays, and other immunological techniques.
Automatically generated - may contain errors
Lab products found in correlation
Propidium iodide, by Thermo Fisher Scientific (3 mentions)
Lps from e coli o111 b4, by Merck Group (1 mentions)
Anti cd103 m290, by BioXCell (1 mentions)
Anti tcr γδ uc7 13d5, by BioXCell (1 mentions)
Armenian hamster igg, by BioXCell (1 mentions)
Carboxyfluorescein succinimidyl ester (cfse), by Thermo Fisher Scientific (1 mentions)
2 protocols using rat igg2a rtk2758
1
Quantifying Epithelial Shedding in Sepsis
2
Analyzing Phagocytosis of Apoptotic Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
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For phagocytosis analysis, apoptotic GL261 cells were incubated with 1 μl/ml carboxyfluorescein succinimidyl ester (CFSE) (Thermo Fisher Scientific) in PBS for 20 min at 37 °C. BMDMs were co-cultured with labeled GL261 or red fluorescent-conjugated latex beads (Sigma-Aldrich). To block CD169, BMDMs were incubated with 2 μg/ml of anti-CD169 antibody (3D6.112) (Bio-Rad) or rat IgG2a (RTK2758) (Biolegend) for 30 min at 37 °C before co-culture. Phagocytosis was analyzed by flow cytometry.
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