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Ebio1d4b

Manufactured by Thermo Fisher Scientific
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The EBio1D4B is a laboratory instrument designed for electrophoresis applications. It provides a controlled and consistent environment for separating and analyzing biological molecules, such as proteins or nucleic acids.

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Lab products found in correlation

Live dead staining, by Thermo Fisher Scientific (1 mentions) Anti cd107a, by Thermo Fisher Scientific (1 mentions) Brefeldin a monensin, by Thermo Fisher Scientific (1 mentions) Facscanto 2, by BD (1 mentions) Monensin and brefeldin a, by Thermo Fisher Scientific (1 mentions) Ionomycin, by Merck Group (1 mentions) Mp6 xt22, by Thermo Fisher Scientific (1 mentions) Mp6 xt22, by BioLegend (1 mentions)

3 protocols using ebio1d4b

1

ILC Function Assay Protocol

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For ILC function assays, spleen cells were stimulated for 4 h with plate bound anti-NK1.1 in the presence of 1 × protein transport inhibitor cocktail (PTIC) containing Brefeldin A/Monensin (eBioscience, Thermo Fisher Scientific) and anti-CD107a (eBio1D4B, eBioscience, Thermo Fisher Scientific) in complete Iscove's DMEM medium (Corning). Cells were incubated during stimulation at 37°C in 5% CO2 for 4 h. Cells were then first surface stained then intracellularly stained to measure function. ILC phenotypes were measured directly ex vivo. Spleen cells were stained following procedures indicated below after fixable Live/Dead staining (Invitrogen).
Ivanova D.L., Krempels R., Denton S.L., Fettel K.D., Saltz G.M., Rach D., Fatima R., Mundhenke T., Materi J., Dunay I.R, & Gigley J.P. (2020). NK Cells Negatively Regulate CD8 T Cells to Promote Immune Exhaustion and Chronic Toxoplasma gondii Infection. Frontiers in Cellular and Infection Microbiology, 10, 313.
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2

CD8+ T Cell Degranulation Assay

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Following degranulation, CD8+ T cells transiently express cell surface CD107a/b (33 (link)). In order to detect alloantigen-induced CD8+ T cell degranulation, spleen cells from transplant recipient mice were isolated in single-cell suspension and co- cultured in 96-well cell culture-treated round-bottom plates in 1:1 ratio (200,000:200,000) with T-cell-depleted spleen cells from donor-origin mice in complete RPMI in the presence of eFluor 660-conjugated CD107a (eBio1D4B), and CD107b (eBioABL-93, (eBioscience, Affymetrix, Santa Clara, CA) followed by a 4 hour incubation (37°C, 5% CO2). Positive control wells were additionally treated with phorbol 12-myristate 13-acetate (PMA, 10ug/ml) and ionomycin (100ug/ml) during the 4 hour incubation. Cells were then incubated with fluorophore-conjugated antibodies and analyzed by flow cytometry.
Mathern D.R., Horwitz J, & Heeger P.S. (2019). Absence of recipient C3aR1 signaling limits expansion and differentiation of alloreactive CD8+ T cell immunity and prolongs murine cardiac allograft survival. American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons, 19(6), 1628-1640.
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3

Spleen Cell Cytokine and Cytotoxicity Assay

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Spleen cell suspensions were cultured during 5 h with medium, 5 μg/ml TSKB20 (ANYKFTLV) peptide (Genscript Inc.), 50 nM PMA plus 0.5 μg/ml ionomycin (Sigma-Aldrich) in the presence of Monensin and Brefeldin A (eBioscience). When indicated a PE-labeled anti-CD107a mAb (eBioscience, eBio1D4B) was included during the culture period. After culture, the cells were surface stained, fixed and permeabilized with BD Cytofix/Cytoperm and Perm/Wash (BD Biosciences) according manufacturer's instruction. Cells were incubated with FITC-labeled antibodies to IFNγ (eBioscience, XMG1.2) or Perforin-1 (eBoscience, eBioOMAK-D), PerCP/PerCP-eFluor 710 labeled antibody to TNF (Biolegend, MP6-XT22) or Granzyme A (eBiosciences, GzA-3G8.5) and/or APC/AF647-labeled antibody to TNF (Biolegend, MP6-XT22) or Granzyme B (Biolegend, GB11). Stained cells were acquired on FACSCanto II (BD Biosciences).
Tosello Boari J., Araujo Furlan C.L., Fiocca Vernengo F., Rodriguez C., Ramello M.C., Amezcua Vesely M.C., Gorosito Serrán M., Nuñez N.G., Richer W., Piaggio E., Montes C.L., Gruppi A, & Acosta Rodríguez E.V. (2018). IL-17RA-Signaling Modulates CD8+ T Cell Survival and Exhaustion During Trypanosoma cruzi Infection. Frontiers in Immunology, 9, 2347.
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