The fluorescence-labeled monoclonal antibodies were as follows: FITC-anti-CD3, PerCp-Cy5.5-anti-CD8, APC-Cy7-anti-CD11b, PerCp-Cy5.5-anti-F4/80, APC-anti-CD4, and PE-Cy7-anti-CD45 were purchased from Biolegend (San Diego, CA, USA); FITC-anti-CD11b was from BD Biosciences (San Jose, CA, USA); and PerCp-Cy5.5-anti-F4/80 was from eBioscience (San Diego, CA, USA). MNCs were stained with the indicated antibodies as previously described37 (link). Data were collected on a FACSCalibur flow cytometer (BD Biosciences) and analyzed using FlowJo 7.6.1 (Tree Star, Ashland, OR).
Percp cy5.5 anti cd8
Manufactured by BioLegend
Sourced in United States
PerCP-Cy5.5-anti-CD8 is a fluorescent-conjugated antibody that recognizes the CD8 cell surface marker. It is used for the identification and analysis of CD8+ T cells in flow cytometry applications.
Automatically generated - may contain errors
Lab products found in correlation
Anti cd4 pe cy7, by Thermo Fisher Scientific (2 mentions)
Anti cd45 fitc, by BioLegend (2 mentions)
Trustainfcx anti mouse cd16 32 fcr block, by BioLegend (2 mentions)
Anti b220 pe, by BioLegend (2 mentions)
Anti mhcii apc, by Thermo Fisher Scientific (2 mentions)
Apc efluor780 anti cd11b, by Thermo Fisher Scientific (2 mentions)
Cytexpert software, by Beckman Coulter (2 mentions)
Anti cd3 fitc, by BioLegend (1 mentions)
Anti cd4 apc, by BioLegend (1 mentions)
Anti cd45 pe cy7, by BioLegend (1 mentions)
Anti cd11b fitc, by BD (1 mentions)
Anti cd11b apc cy7, by BioLegend (1 mentions)
Percp cy5.5 anti f4 80, by BioLegend (1 mentions)
Facscalibur flow cytometer, by BD (1 mentions)
Cytoflex s flow cytometer, by Beckman Coulter (1 mentions)
Anti b220 pe, by BD (1 mentions)
Anti cd4 pe, by BD (1 mentions)
Anti cd45 pacific blue, by BioLegend (1 mentions)
Facscanto 2, by BD (1 mentions)
Anti f4 80 apc, by BioLegend (1 mentions)
4 protocols using percp cy5.5 anti cd8
1
Multicolor Flow Cytometry Analysis
2
Multi-Parametric Flow Cytometry Panel
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were resuspended in 100 μl FCB, blocked with 0.5 μl TruStainFcX (anti-mouse CD16/32 FcR block, Biolegend) for 5 min at room temperature, and stained for 30 min at 4 °C with: FITC-anti-CD45 (1:1000, Biolegend), PE-Cy7-anti-CD4 (1:200, eBioscience), PerCP-Cy5.5-anti-CD8 (1:200, Biolegend), PE-anti-B220 (1:200, Biolegend), APC-anti-MHCII (1:200, eBioscience), and APC-eFluor780-anti-CD11b (1:200, eBioscience) (Supplemental Table 1 ). Cells were then fixed in 1% PFA for 1 h. Finally, cells were resuspended in 500 μl FCB, labelled with DAPI to exclude cell debris, and analyzed with a CytoFLEX S flow cytometer (Beckman-Coulter) equipped with the CytExpert software (Beckman-Coulter).
3
Cecal Immune Cell Profiling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cecal patch was isolated and washed with PBS, then gently ground using the plunger of a 5 mL syringe. The cell suspension was passed through a 70 μm cell strainer and collected for flow cytometry after centrifugation. The cells were incubated with Pacific Blue anti-CD45 (BioLegend, San Diego, CA, USA), PE anti-CD4 (BD Biosciences, San Jose, CA, USA), PerCP-Cy5.5 anti-CD8 (BioLegend), APC anti-F4/80 (BioLegend), PE anti-B220 (BD Biosciences) mAbs for 30 min at 4 °C. The samples were washed twice with PBS, then acquired using BD FACS Canto II and analyzed with FlowJo v10.
4
Flow Cytometric Analysis of Immune Cells in EAE
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were resuspended in 100 ml FCB, blocked with 0.5 ml TruStainFcX (anti-mouse CD16/32 FcR block, Biolegend) for 5 min at room temperature, and stained for 30 min at 4°C with: FITC-anti-CD45 (1:1000, Biolegend), PE-Cy7-anti-CD4 (1:200, eBioscience), PerCP-Cy5.5-anti-CD8 (1:200, Biolegend), PE-anti-B220 (1:200, Biolegend), APC-anti-MHCII (1:200, eBioscience), and APC-eFluor780-anti-CD11b (1:200, eBioscience) (Supplemental Table 1). Cells were then xed in 1% PFA for 1 h. Finally, cells were resuspended in 500 ml FCB, labelled with DAPI to exclude cell debris and analyzed with a CytoFLEX S ow cytometer (Beckman-Coulter) equipped with CytExpert software (Beckman-Coulter).
Quanti cation of IC100 in Tissues. IC100 was quanti ed in brain, spinal cord, liver and spleen at 35 days post-induction (dpi) of EAE using a proprietary assay developed by In amaCORE, LLC using Meso Scale Technology. Protein lysates were obtained as described in [29] . The assay was read using the QuickPlex SQ 120 instrument (Meso Scale Diagnostics, Maryland).
Quanti cation of IC100 in Tissues. IC100 was quanti ed in brain, spinal cord, liver and spleen at 35 days post-induction (dpi) of EAE using a proprietary assay developed by In amaCORE, LLC using Meso Scale Technology. Protein lysates were obtained as described in [29] . The assay was read using the QuickPlex SQ 120 instrument (Meso Scale Diagnostics, Maryland).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!