Lymphocytes were isolated as described and stained with anti-mouse IgD PerCP-Cy5.5 (1:400, 405710), LPAM (integrin α4β7) PE (1:100, 120605; (Biolegend, San Diego, CA), B220 V500 (1:400, 561227), CD19 APC-H7 (1:200, 560143), CD80 PE (1:500, 553769), CD273 APC (1:200, 560086), CD138 PE (1:200, 553714), IgM PE-Cy7 (1:200, 552867; BD Biosciences), CCR9 PE-Cy7 (1:100, 25-1991), CD73 PE-Cy7 (1:50, 25-0731), IgA PE (1:50, 12-4204), GL7 eFluor 450 (1:100, 48-5902), CD38 Alexa700 (1:800, 56-0381), CD21/35 Pacific Blue (1:800, 57-0212; eBiosciences) or CCR10 APC (1:100, FAB2815A; R and D systems. Minneapolis, MN) and were analysed using LSR II (BD Biosciences) or Navios (Beckman Coulter, Brea, CA) flow cytometers. For sorting, cells were labelled with anti-mouse CD138 PE (1:200, 553714), CD19 PE-Cy7 (1:200, 552854), CD80 APC (1:200, 560016) and GL7 FITC (1:100, 553666) before sorting using a FACSAria (BD Biosciences). Cells were sorted into tubes that had been coated with 2% BSA/PBS overnight, and pelleted by centrifugation at 600 g before being resuspended in PBS and injected into recipient mice or used for gene sequence analysis.
Cd73 pe cy7
Manufactured by Thermo Fisher Scientific
Sourced in United States
CD73 PE-Cy7 is a fluorescently-labeled antibody used for the detection and analysis of CD73-positive cells in flow cytometry applications. CD73 is an enzyme involved in the regulation of adenosine signaling pathways.
Automatically generated - may contain errors
Lab products found in correlation
Facsaria, by BD (1 mentions)
Cd80 pe, by BD (1 mentions)
Cd19 pe cy7, by BD (1 mentions)
Cd138 pe, by BD (1 mentions)
Ccr10 apc, by R&D Systems (1 mentions)
Cd19 apc h7, by BD (1 mentions)
Cd80 apc, by BD (1 mentions)
Gl7 fitc, by BD (1 mentions)
Igd percp cy5, by BioLegend (1 mentions)
Navios, by Beckman Coulter (1 mentions)
Cd273 apc, by BD (1 mentions)
Igm pe cy7, by BD (1 mentions)
Iga pe, by Thermo Fisher Scientific (1 mentions)
Tgf β1, by Thermo Fisher Scientific (1 mentions)
Recombinant mouse il 2, by Thermo Fisher Scientific (1 mentions)
Cd25 pe, by Thermo Fisher Scientific (1 mentions)
Il 17 pe, by Thermo Fisher Scientific (1 mentions)
Foxp3 pe cy7, by Thermo Fisher Scientific (1 mentions)
Cd4 pe cy7, by BioLegend (1 mentions)
Apc il 17, by Thermo Fisher Scientific (1 mentions)
3 protocols using cd73 pe cy7
1
Lymphocyte Isolation and Characterization
2
Isolation and Analysis of T Cell Subsets
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The following antibodies from eBioscience (San Diego, CA, USA) were used: anti-mouse CD4 APC-H7 (GK1.5), CD25 PE (PC61.5), IL-17 PE (eBio17B7), IL-17 APC (eBio17B7), Foxp3 PE-Cy7 (FJK-16s), IgG 2a PE-Cy7 (eBR2a), CD39 PE (24DMS1), CD73 PE-Cy7 (Ty/11.8), CTLA-4 PE (UC10-4F10-11), Lag-3 APC (C9b7w), CCR9 PE (eBioCW-1.2), α4β7 PE (DATK32), B220 (RA3-6b2), CD19 APCH-7 (1D3), CD11c (N418), CD62L FITC (MEL-14), CD45.1 PE-cy7 (A20) purified anti-CD3 (145-2C11), and purified CD16/32 (93). From Biolegend (San Diego, CA, USA), we used CD4 PECy7 (GK 1.5), I-Ab FITC (25-9-17) and CD11b APC (M1/70), and purified α-IFNγ (XMG1.2). Recombinant mouse IL-2, TGF-β1, IL-6, and IL-1β were purchased from eBioscience. All-trans-retinoic acid, OVA protein, PMA, and ionomycin were purchased from Sigma Aldrich (St. Louis, MO, USA).
3
Adoptive Transfer of Antigen-Specific B Cells
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Cells from spleens and lymph nodes were prepared as described (8) . Red blood cells were lysed by ACK lysing buffer (Gibco). Cell suspensions were blocked by CD16/32 (93, eBioscience) diluted in FACS buffer (PBS supplemented with 0.5% BSA plus 2mM EDTA), followed with staining cocktail. PD-L2 biotin (TY25) and B220-BV421 or BV510 (RA3-6B2) and Str. BV421 or BV711 were from BioLegend, NP was conjugated in house with PE for detected antigen specific B cells (8) . CD38 APC (90), GL7 eFluor 450 (GL7), CD86 PE-Cy5 (GL1), CD80 PE-Cy5(16-10A1), and CD73 PE-Cy7 (eBioTY) were from eBioscience. CXCR4 biotin (2B11), Fas PE-Cy7 (Jo2) and CD138 APC or BV421 (281-2) were from BD Bioscience. Samples were analyzed using BD LSRFortessa Analyzer (BD Biosciences, USA) with the software BD FACSDiva (BD Biosciences). Data were analyzed offline with FlowJo (FlowJo LLC, USA).
For adoptive transfer experiment, 2x10 5 NP + B220 + cells from spleens of fluorescent protein labelled QM background mice were transferred into C57BL6/J hosts 1 d before immunization with NP-CGG in alum on rear feet. In co-transfer experiments, a mix of 1x10 5 of NP + B220 + B cells of each genotype respectively were injected i.v.
For adoptive transfer experiment, 2x10 5 NP + B220 + cells from spleens of fluorescent protein labelled QM background mice were transferred into C57BL6/J hosts 1 d before immunization with NP-CGG in alum on rear feet. In co-transfer experiments, a mix of 1x10 5 of NP + B220 + B cells of each genotype respectively were injected i.v.
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