Rat igg1 k isotype control clone ebrg1

Manufactured by Thermo Fisher Scientific

Rat IgG1 k isotype control (clone eBRG1) is a laboratory reagent used as a control for flow cytometry and other immunoassay applications. It is an antibody of the IgG1 subclass and kappa light chain isotype derived from a rat.

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Lab products found in correlation

Anti fitc macs beads 130 048 701, by Miltenyi Biotec (2 mentions) Elisa pierce tmb substrate kit, by Thermo Fisher Scientific (2 mentions) Dulbecco s pbs, by Avantor (2 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) Glutamax, by Thermo Fisher Scientific (1 mentions) L glutamine, by Thermo Fisher Scientific (1 mentions) Rhil 2, by Novartis (1 mentions) Proleukin, by Novartis (1 mentions)

Close spelling variants of this product

Rat IgG1 isotype control (8 citations) IgG1 isotype control (2 citations) Control rat IgG1 (3 citations) IgG1 K (3 citations) Isotype controls (54 citations) Rat IgG1 (6 citations) EBRG1 (4 citations)

3 protocols using rat igg1 k isotype control clone ebrg1

Immunoblotting and FACS Antibody Validation

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Anti-ISG15 rabbit serum, gift of Klaus-Peter Knobeloch, was used as primary antibody for immunoblotting. Secondary antibodies for immunoblotting were from KCL: goat anti-rabbit HRP (474-1506) and goat anti-mouse HRP (474-1806), used for the IgG heavy chain blot. Fluorescently labelled antibodies for cytofluorometry (FACS) against Ly6C (clone HK1.4), CD11b (clone M1/70), I-A/I-E (clone M5/114.15.2), CD11c (clone N418), Ly6G (clone 1A8), F4/80 (clone BM8), CD103 (clone 2E7), CD8α (clone 53-6.7), CD45R/B220 (clone RA3-6B2), CD49b (clone DX5), TCRβ (clone H57-597) and purified CD16/32 (anti Fc-γ receptor III/II, clone 93) were purchased from Biolegend. Anti-mouse IL-1β (clone NJTEN3), anti-mouse IL-12 (clone C15.6) and Rat IgG1 k isotype control (clone eBRG1) were purchased from eBioscience. Anti-FITC MACS beads (130-048-701) used for negative selection for the intracellular staining were from Miltenyi Biotec. ELISA Pierce TMB substrate kit (34021) was from Thermo Fisher Scientific. Dulbecco’s PBS was from VWR (21-031-CV).

Napolitano A., van der Veen A.G., Bunyan M., Borg A., Frith D., Howell S., Kjaer S., Beling A., Snijders A.P., Knobeloch K.P, & Frickel E.M. (2018). Cysteine-Reactive Free ISG15 Generates IL-1β–Producing CD8α+ Dendritic Cells at the Site of Infection. The Journal of Immunology Author Choice, 201(2), 604-614.

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Immunoblotting and Flow Cytometry Analyses

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Anti-ISG15 rabbit serum, a gift from K.-P. Knobeloch, was used as primary Ab for immunoblotting. Secondary Abs for immunoblotting were from KPL: goat anti-rabbit HRP (474-1506) and goat anti-mouse HRP (474-1806), used for the IgG H chain blot. Fluorescently labeled Abs for FACS against Ly6C (clone HK1.4), CD11b (clone M1/70), I-A/I-E (clone M5/114.15.2), CD11c (clone N418), Ly6G (clone 1A8), F4/80 (clone BM8), CD103 (clone 2E7), CD8α (clone 53-6.7), CD45R/B220 (clone RA3-6B2), CD49b (clone DX5), TCRβ (clone H57-597), and purified CD16/32 (anti Fc-γ receptor III/II, clone 93) were purchased from BioLegend. Anti-mouse IL-1β (clone NJTEN3), anti-mouse IL-12 (clone C15.6), and Rat IgG1 k isotype control (clone eBRG1) were purchased from eBioscience. Anti-FITC MACS beads (130-048-701) used for negative selection for the intracellular staining were from Miltenyi Biotec. ELISA Pierce TMB substrate kit (34021) was from Thermo Fisher Scientific. Dulbecco’s PBS was from VWR (21-031-CV).

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Modulation of Organoid Growth by iNKT Cells

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HeLa cells with or without stable overexpression of hCD1D were grown in DMEM supplemented with 10% heat-inactivated bovine calf serum, 1X Glutamax, 1X penicillin/streptomycin (all from Life Technologies) and incubated with 100 ng/ml α-GC for 24 h. Subsequently, iNKT DN32.D3 hybridoma cells (Bendelac et al., 1995 (link)) were incubated with HeLa and HeLa-CD1D cells for another 24 h in serum-free advanced DMEM/F12 medium supplemented with 1X l-Glutamine, 1X penicillin/streptomycin (Life Technologies), and rhIL-2 (10 U/ml; Proleukin; Novartis). SNs were centrifuged for cell depletion and included into the regular organoid medium as 40% (vol/vol). IFN-γ levels were measured with a commercially available Elisa kit from BD. A standard curve was recorded using recombinant mouse IFN-γ protein. In some experiments, SNs were treated with functional grade anti–mouse IFN-γ monoclonal (clone XMG1.2) or rat IgG1 K isotype control (clone eBRG1; both from eBioscience; final concentrations 0.5 µg/ml) 30 min before addition to the organoids.

Farin H.F., Karthaus W.R., Kujala P., Rakhshandehroo M., Schwank G., Vries R.G., Kalkhoven E., Nieuwenhuis E.E, & Clevers H. (2014). Paneth cell extrusion and release of antimicrobial products is directly controlled by immune cell–derived IFN-γ. The Journal of Experimental Medicine, 211(7), 1393-1405.

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