Cd73 ty 11.8

Manufactured by BioLegend

CD73 (TY/11.8) is a monoclonal antibody that recognizes the CD73 cell surface molecule. CD73 is an enzyme that catalyzes the conversion of extracellular adenosine monophosphate (AMP) to adenosine.

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Lab products found in correlation

Streptavidin microbeads, by Miltenyi Biotec (1 mentions) Facscanto, by BD (1 mentions) Facscalibur, by BD (1 mentions) Duoset elisa development system, by R&D Systems (1 mentions) Accutase, by Merck Group (1 mentions) Ifn γ, by Thermo Fisher Scientific (1 mentions) Cd11b m1 70, by BioLegend (1 mentions) Il 17a tc11 18h10.1, by BioLegend (1 mentions) Pna fitc, by Vector Laboratories (1 mentions) Cd69 h1.2f3, by BioLegend (1 mentions) Cd19 1d3, by BD (1 mentions) Horseradish peroxidase conjugated anti rabbit igg, by Cell Signaling Technology (1 mentions) Cd21 7e9, by BioLegend (1 mentions) Cd25 pc61, by BioLegend (1 mentions) Cd95 jo2, by BD (1 mentions)

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Anti-CD73 (TY/11.8) (2 citations)

3 protocols using cd73 ty 11.8

Purification and Transfer of Splenic Immune Cells

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Splenic B-lymphocytes and T-cells were purified by negative depletion of CD11b⁺, CD11c⁺, TER-119⁺, CD3ε⁺ populations and in some experiments CD73⁺ (for B-lymphocyte purification) or CD11b⁺, CD11c⁺, TER-119⁺ and B220⁺ cells (for T-cell purification), using biotin-conjugated antibodies and streptavidin microbeads (Miltenyi). NOD-scid recipients were intravenously (i.v) injected with indicated B-lymphocytes and/or T-cells and monitored for T1D development. Specific biotinylated antibodies used from BD Biosciences were: CD11b (M1/70), CD11c (HL3), CD3ε (145-2C11), TER-119 (TER119), B220 (RA3-6B2). CD73 (TY/11.8) was obtained from BioLegend.

Ratiu J.J., Racine J.J., Hasham M.G., Wang Q., Branca J.A., Chapman H.D., Zhu J., Donghia N., Philip V., Schott W.H., Wasserfall C., Atkinson M.A., Mills K.D., Leeth C.M, & Serreze D.V. (2017). Genetic and small molecule disruption of the AID/RAD51 axis similarly protect NOD mice from type 1 diabetes through expansion of regulatory B-lymphocytes,. Journal of immunology (Baltimore, Md. : 1950), 198(11), 4255-4267.

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Characterization of CD4+ T Cells and Astrocytes

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After 48 h or 96 h of incubation, non-adherent CD4 T cells were removed by washing the co-culture 3 times with PBS (2% FBS). The remaining adherent cells (T cells and astrocytes) were incubated with accutase (Millipore) at 37°C for 40 min, collected and analyzed by flow cytometry. CD4⁺ cells were stained with the following mouse specific monoclonal antibodies: CD3 (clone 17A2), CD4 (GK1.5), MHCII (M5/114.15.2), CD39 (24DMS1), Rorg (gamma, greek letter) t (lower case) (AFKJS-9), IFN-γ (XMG1.2), CD25 (PC61 5), FoxP3 (FJK-165), CD44 (IM7), CD62L (MEL-14) (eBioscience), CD73 (TY11.8) and IL-17A (TC11-18H10.1) (Biolegend). Astrocytes were stained for CD11b (M1/70) (Biolegend), VCAM-1 (429) (BD Bioscience), ICAM-1 (YN1/1.7.4) (eBioscience). All samples were acquired with a FACSCalibur or FACSCanto (Becton Dickinson) and analyzed with FlowJo software (FlowJo v10.0.7). IL-6 protein concentration was determined using the DuoSet ELISA Development System according to the manufacturer's instructions (R&D Systems).

Filipello F., Pozzi D., Proietti M., Romagnani A., Mazzitelli S., Matteoli M., Verderio C, & Grassi F. (2016). Ectonucleotidase activity and immunosuppression in astrocyte-CD4 T cell bidirectional signaling. Oncotarget, 7(5), 5143-5156.

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Comprehensive Mouse Antibody Panel for Flow Cytometry

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Anti-mouse antibodies used for flow cytometry include the following: CD95(Jo2), CD138(281–2), CD4(RM4–5), CD38(90), MHC-II(2G9), CD23(B3B4), CD80(B7–1), CD44(IM7), B220(RA3–6B2), CD11b(M1/70), IgM(R6–60.2), IgD(11–26c.2a), CD19(1D3) and Mouse BD Fc block (2.4G2) from BD Biosciences. CD8α(53–6.7), GL7(GL7), CD86 (GL1), CD21(7E9), CD69(H1.2F3), CD25(PC61) and CD73 (Ty/11.8) were from Biolegend. Anti-NFκ-B (D14E12), anti-LSD1 (C69G12) and horseradish peroxidase conjugated anti-rabbit IgG were from Cell Signaling Technology. Alkaline phosphatase conjugated anti-mouse IgG-AP, alkaline phosphatase conjugated anti-mouse IgG2c-AP and anti-mouse IgG(H+L) were from Southern Biotech. PNA-FITC was from Vector laboratories and R848 was from Invivogen.

Acharya M., Raso F., Sagadiev S., Gilbertson E., Kadavy L., Li Q.Z., Yan M., Stuart L.M., Hamerman J.A, & Lacy-Hulbert A. (2020). B Cell αv Integrins Regulate TLR-Driven Autoimmunity. The Journal of Immunology Author Choice, 205(7), 1810-1818.

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