Spleen cell surface antigens were analyzed by use of antibodies: Anti-CD3-PE clone 145-2C11, anti-CD4-APC clone RM 4-5, anti-CD8-APC clone 53-6.7, anti-CD11b-PECy7 clone M1/70, anti-Ly6G(Gr-1)-APC clone RB6-8C5, and anti-F4/80-FITC clone BM8 (all eBioscience, San Diego, CA) after blocking of FC antibody-binding by anti-CD16/CD32 (BD Biosciences, Franklin Lakes, NJ) for 10 min. Cells were fixed in PBS with 2% methanol-free formaldehyde and analyzed within 1–3 days on a FACScanto flow cytometer (BD Biosciences). Data analyses and layouts were performed using FlowJo V10 (Tree Star, Ashland, OR).
Anti cd8 apc clone 53 6
Manufactured by Thermo Fisher Scientific
The Anti-CD8-APC clone 53-6.7 is a fluorescently-labeled monoclonal antibody that binds to the CD8 molecule expressed on the surface of T cells. It is designed for use in flow cytometry applications to identify and quantify CD8+ T cells within a cell population.
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Lab products found in correlation
Pe cy7 anti cd11b clone m1 70, by Thermo Fisher Scientific (1 mentions)
Anti cd16 cd32, by BD (1 mentions)
Facscanto flow cytometer, by BD (1 mentions)
Anti f4 80 fitc clone bm8, by Thermo Fisher Scientific (1 mentions)
Anti cd3 pe clone 145 2c11, by Thermo Fisher Scientific (1 mentions)
Oct compound, by Sakura Finetek (1 mentions)
Facsaria flow cytometer, by BD (1 mentions)
Cfse labeled, by Thermo Fisher Scientific (1 mentions)
Anti cd4 bv421 clone gk1.5, by BioLegend (1 mentions)
2 protocols using anti cd8 apc clone 53 6
1
Immune Cell Profiling by Flow Cytometry
2
In Vivo CD4 T Cell Tracking
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Mouse (C57BL/6; The Jackson Laboratory) CD4 T cells from spleen and lymph nodes were prepared (CD4 enrichment kit; STEMCELL Technologies) and CFSE labeled (Molecular Probes). Recipient mice were given vehicle or 30 mg/kg AM095 by gavage. 2 h later, 106 labeled CD4 T cells were adoptively transferred i.v. 16 h later, spleen or lymph nodes were collected and stained with anti-CD4-BV421 (clone GK1.5; BioLegend) and anti–CD8-APC (clone 53–6.7; eBioscience). The samples were analyzed with FACSAria flow cytometer (BD Biosciences). Data were analyzed with FlowJo software v 8.8.7 (Tree Star). Values are expressed as the percentage of CSFE+ CD4+ cells out of total CD4+ cells.
Spleen and lymph nodes were collected from above recipient mice and immediately submerged in OCT compound (Sakura Finetek). Tissues in OCT were quickly frozen using dry ice and then kept at −80°C for long-term storage. Sections of the lymph nodes were stained as above and imaged by confocal microscopy.
Spleen and lymph nodes were collected from above recipient mice and immediately submerged in OCT compound (Sakura Finetek). Tissues in OCT were quickly frozen using dry ice and then kept at −80°C for long-term storage. Sections of the lymph nodes were stained as above and imaged by confocal microscopy.
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