Cell pellets obtained from the striatum and midbrain were resuspended in 100 µL of CB and incubated for 5 min at RT with Zombie NIR Dye (BioLegend, San Diego, CA, USA) to assess cell viability (50 µL/sample; 1:2000 dilution in PBS). After quenching with CB, cells were centrifuged at 2000 rpm for 1 min, and labeled with BV510 anti-CD11b (1:500, M1/70, BioLegend), BV421 anti-CD45 (1:1000, 30F11, BioLegend), PE-Cy7 anti-CD8a (1:400, 53 − 6.7, BioLegend) or BUV395 anti-CD8a (1:200, 53 − 6.7, BD Bioscience), and FITC anti-CD4 (1:1000, GK1.5, BioLegend) or BV711 anti-CD4 (1:200, GK1.5, BioLegend) diluted in CB and the FcR Blocking Reagent (1:50; Miltenyi Biotec, Bergisch Gladbach, Germany) during 15 min at 4 °C in the dark. For intracellular staining, cells were incubated with fixation/permeabilization solution (Invitrogen) during 7 min at 4 °C in the dark and washed with PermWASH solution (Invitrogen) and incubated with FITC anti-Ki67 (1:500, 11F6, BioLegend) diluted in PermWASH solution during 15 min at 4 °C in the dark. Once labeled, samples were centrifuged, resuspended in CB and data was acquired using a CytoFLEX LX Flow Cytometer (Beckman Coulter, Brea, CA). The analysis was performed using the CytExpert 2.3 (Beckman Coulter) and the FlowJo 10.0.7r2 (BD Biosciences, Franklin Lakes, NJ) softwares.
Pe cy7 anti cd8a
Manufactured by BioLegend
Sourced in Germany, United States
PE-Cy7 anti-CD8a is a fluorescent-conjugated antibody that binds to the CD8a receptor on the surface of T cells. It can be used for the identification and analysis of CD8+ T cells in flow cytometry applications.
Automatically generated - may contain errors
Lab products found in correlation
Zombie nir dye, by BioLegend (1 mentions)
Anti cd11b bv510, by BioLegend (1 mentions)
Anti cd45 bv421, by BioLegend (1 mentions)
Anti cd4 bv711, by BioLegend (1 mentions)
Fixation permeabilization solution, by Thermo Fisher Scientific (1 mentions)
Perm wash solution, by Thermo Fisher Scientific (1 mentions)
Cytexpert 2, by Beckman Coulter (1 mentions)
Perm wash solution, by BioLegend (1 mentions)
Anti ki67 fitc, by BioLegend (1 mentions)
Cytoflex lx flow cytometer, by Beckman Coulter (1 mentions)
Fcr blocking reagent, by Miltenyi Biotec (1 mentions)
Anti cd4 fitc, by BioLegend (1 mentions)
Penicillin streptomycin pen strep, by Thermo Fisher Scientific (1 mentions)
Anti cd11b bv650, by BioLegend (1 mentions)
Pecy5 anti cd4, by BioLegend (1 mentions)
Liberase, by Thermo Fisher Scientific (1 mentions)
Anti cd45 apc cy7, by BioLegend (1 mentions)
Anti ly6c pe, by BioLegend (1 mentions)
Anti cd11c bv421, by BioLegend (1 mentions)
Anti cd206 apc, by BioLegend (1 mentions)
2 protocols using pe cy7 anti cd8a
1
Multicolor Flow Cytometry for Cell Viability
2
Nanoparticle-mediated Cancer Immunotherapy Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
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DOTAP (1,2-dioleoyl-3-trimethylammonium-propane) (890890) and cholesterol (700000) lipids were purchased from Avanti Polar Lipids, Inc., Alabaster, AL, USA. Nanoplasmid (NTC 9385R-MCS) containing the CRT gene was purchased from Aldevron (Lincoln, NE, USA). Chloroform (C2432) was purchased from Sigma-Aldrich, St. Louis, MO, USA. HEPES, sterile 1 M buffer (J848), DMEM (11965092), Fetal bovine serum, FBS (10082147), Penicillin-Streptomycin - PenStrep (15140122), PBS (10010023), Collagenase IV (17104019), were purchased from ThermoFisher/Gibco, Waltham, MA, USA. The following Fluorochrome-conjugated monoclonal antibodies (mAbs) for flow cytometry were purchased from BioLegend, San Diego, CA, USA and were listed: APC-Cy7 anti-CD45 (103115), PerCP anti-CD3 epsilon (100325), PE-Cy5 anti-CD4 (100410), PE-Cy7 anti-CD8a (100721), PE anti-Granzyme B (372208), BV510 anti-INFg (505841), BV510 anti-Ly-6G (127633), PE anti-Ly-6C (128007), BV650 anti-CD11b (101239), APC anti-CD206 (141707), BV421 anti-CD11c (117329). AF488 anti-iNOS (53-5920-82) was procured from eBiosciences/ThermoFisher, Waltham, MA, USA. Liberase (5401119001) was procured from Life Technologies, NY, USA. Transcription factor buffer set (562574) was purchased from BD Biosciences, San Jose, CA, USA. B16F10 was provided by Dr. Mary Jo Turk, Dartmouth. Aged C57BL/6 mice were obtained from NIA, Bethesda, MD, USA.
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