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Clone ebr2a

Manufactured by Thermo Fisher Scientific
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The Clone eBR2a is a laboratory equipment designed for specific scientific applications. It serves a core function within the experimental workflow. Further details on the intended use or performance specifications are not available.

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Lab products found in correlation

Intracellular staining permeabilization wash buffer, by BioLegend (1 mentions) Fixable viability dye efluor 506, by Thermo Fisher Scientific (1 mentions) Facscanto 2, by BD (1 mentions) Clone ltf 2, by BioXCell (1 mentions) Cd8 clone yts 169, by BioXCell (1 mentions) Cd4 clone gk1, by BioXCell (1 mentions) Clone il 22jop, by Thermo Fisher Scientific (1 mentions) Anti cd8, by Thermo Fisher Scientific (1 mentions) Cd3 e450, by Thermo Fisher Scientific (1 mentions) Lsrii flow cytometer, by FlowJo (1 mentions) Cd4 fitc, by Thermo Fisher Scientific (1 mentions) Facsdiva software, by BD (1 mentions) Facscanto, by BD (1 mentions)

Close spelling variants of this product

Rat IgG2a isotype antibody (clone eBR2a, Isotype control IgG2aκ (Clone eBR2a) Rat IgG2a kappa isotype control (clone eBR2a; EBR2a

5 protocols using clone ebr2a

1

GILZ Expression in Cultured PBMC

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Cultured PBMC were stained with Fixable Viability Dye eFluor506 (eBioscience) and subsequently fixed with 2% paraformaldehyde. After permeabilization (Intracellular Staining Permeabilization Wash Buffer, BioLegend), cells were stained intracellularly for GILZ (clone CFMKG15, eBioscience) or the corresponding rat IgG2a kappa isotype control (clone eBR2a, eBioscience). Data was acquired on the FACSCantoII (BD Biosciences) and analyzed using FlowJo v10.8 (BD).
Dankers W., Northcott M., Bennett T., D’Cruz A., Sherlock R., Gearing L.J., Hertzog P., Russ B., Miceli I., Scheer S., Fujishiro M., Hayakawa K., Ikeda K., Morand E.F, & Jones S.A. (2022). Type 1 interferon suppresses expression and glucocorticoid induction of glucocorticoid-induced leucine zipper (GILZ). Frontiers in Immunology, 13, 1034880.
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2

Monocyte, Macrophage, and T Cell Depletion

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To deplete monocytes, wildtype C57BL/6 mice received I.V. injection of 200 μL of clodronate liposomes (or PBS control liposomes) 15–18 hours prior to infection [35 (link)]. Anti-CSF1R antibody (2 mg/mL, clone AFS98, eBioscience) was used to deplete bladder-resident macrophages. Animals received two I.V. injections, on consecutive days, of anti-CSF1R antibody or isotype control (clone eBR2a, eBioscience). We administered 400 μg/mouse on day 1 and 200 μg/mouse on day 2, to decrease the impact on circulating monocytes. To deplete T cells, 100 μg of CD4 (clone GK1.5, Bio X Cell) and 100 μg of CD8 (clone YTS 169.4, Bio X Cell) per mouse were injected together intraperitoneally 24 hours prior to primary infection. 200 μg of isotype control (clone LTF-2, Bio X Cell) per mouse was injected intraperitoneally. The depletion was repeated 5 days post-infection, and once a week to maintain the depletion until challenge infection.
Mora-Bau G., Platt A.M., van Rooijen N., Randolph G.J., Albert M.L, & Ingersoll M.A. (2015). Macrophages Subvert Adaptive Immunity to Urinary Tract Infection. PLoS Pathogens, 11(7), e1005044.
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3

DSS-Induced Colitis Model in Mice

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Mouse experiments were performed in strict accordance with the guidelines of the European Community (86/609/EEC) and the French National Committee (87/848) for the care and use of laboratory animals. The study plan was approved by the Institutional Review Board at the Animal Facility of the Institute de Génétique Moléculaire de Montpellier (IGMM) and the Regional Ethics Committee for Animal Experimentation of Languedoc-Roussillon (agreement n° CEEA-LR-12087).
The DSS experiments described in Fig. 1 were performed at Amylgen (In vivo drug discovery, Montferrier sur Lez, France), whereas those described in Figs 2, 3 and 6 were done at the animal facility of IGMM. For DSS-induced colitis, age- and sex-matched C57BL6 mice received one cycle of 2.5% (w/v) DSS administered via drinking water. Mice received 40 mg/kg ABX464 suspended in MC or MC only via gavage when indicated. For antagonizing IL-22 in vivo, mice received either blocking IL-22 antibody (Ebioscience; clone IL-22JOP) or isotype control (Ebioscience; clone eBR2a).
Chebli K., Papon L., Paul C., Garcel A., Campos N., Scherrer D., J. Ehrlich H., Hahne M, & Tazi J. (2017). The Anti-Hiv Candidate Abx464 Dampens Intestinal Inflammation by Triggering Il-22 Production in Activated Macrophages. Scientific Reports, 7, 4860.
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4

CD8+ T-Cell Depletion and Caloric Restriction

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Forty mice were injected with 20 µg of either anti-CD8 (Clone 53–6.7, eBioscience) or isotype control antibodies (Clone eBR2a, eBioscience) via intraperitoneal injection on days 6, 5, and 4 prior to tumor challenge. Ten mice treated with anti-CD8 and ten with isotype control were placed on a caloric restriction diet as described above, while the rest were maintained on an AL diet. Depletion was maintained after tumor challenge for the duration of the experiment with administration of anti-CD8 antibody every 3 days. CD8 T-cell depletion was confirmed via flow cytometry. Splenocytes were stained with CD3 e450 (Clone 17A2, eBioscience), CD4 FITC (Clone RM4–5, eBioscience), and CD8 APC-Cy7 (Clone 53–6.7, Life Technologies). Samples were sorted on a BD Biosciences LSR II flow cytometer and analyzed using FlowJo v10.6.2.
Manukian G., Kivolowitz C., DeAngelis T., Shastri A., Savage J.E., Camphausen K., Rodeck U., Zarif J, & Simone N.L. (2021). Caloric restriction impairs regulatory T-cells within the tumor microenvironment after radiation and primes effector T-cells. International journal of radiation oncology, biology, physics, 110(5), 1341-1349.
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5

Flow Cytometry Analysis of Immune Cell Markers

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Flow data were acquired using BD FACS DIVA software (BD Bioscience, Franklin Lakes, NJ, USA) on FACSCanto (BD bioscience). Flow data were analyzed by Flowjo software vs 10.2. Flow cytometry antibodies used; anti-mouse CD11c PerCp-cyanine5.5, PE; clone N418, MHC classII (I-A/I-E) APC; cloneM5/114.15.2, CD86(B7–2) PE; clone GL1, CD83 FITC; clone Michel-17, CD11b PerCP-Cyanine5.5; clone M1/70, LY-6G(Gr-1) APC; clone1A8-LY6g, CD94 FITC; clone 18d3, F4/80 PerCP-Cyanine5.5; clone BM8, CD68 PE; clone FA-11, CD3e PE; clone 145–2C11, CD4 APC; clone GK1.5, CD8 PerCP-Cyanine 5.5; clone 53–6.7, CD9 APC; clone eBioKMC8 (KMC8), CCR7 APC; clone 4B12. (affymetrix, eBioscience, Thermofisher Scientific, Waltham MA, USA) and Isotype controls; American Hamster IgG isotype control PE, FITC; clone eBio299Arm, Rat IgG2b Isotype control PE; clone eB149/10H5, Rat IgG2b K Isotype control APC; clone eB149/10H5, Rat IgG2a K Isotype control APC, FITC, Percp-cyanine5.5; clone eBR2a (affymetrix, eBioscience, Thermofisher Scientific, Waltham MA, USA). Unconjugated CD11c monoclonal antibody; clone N418 (Thermofisher Scientific, Waltham MA, USA). Rabbit anti Hamster IgG (H+L) secondary antibody TRITC conjugate. (Thermofisher Scientific, Waltham MA, USA)
Elsayed R., Kurago Z., Cutler C.W., Arce R., Gerber J., Celis E., Sultan H., Elashiry M., Meghil M., Sun C., Auersvald C.M., Elsayed R., Elshikh M.E., Isales C, & Elsalanty M.E. (2020). Role of dendritic cell - mediated immune response in oral homeostasis: a new mechanism of osteonecrosis of the jaw. FASEB journal : official publication of the Federation of American Societies for Experimental Biology, 34(2), 2595-2608.
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