Human PBMCs at 106/tube were stained in duplicate with allophycocyanin (APC)-H7-anti-CD3, BV510-anti-CD4, fluorescein isothiocyanate (FITC)-anti-CD45RA, phycoerythrin (PE)-Cy™7-anti-CCR7, peridinin-chlorophyll proteins (PerCP)-Cy™5.5-anti-CXCR5, PE-anti-ICOS, BV421-anti-PD-1, PE-CF594-anti-CD154, or proper IgG isotype controls (Becton Dickinson, San Diego, CA, USA) at room temperature for 30 minutes. After being washed with phosphate-buffered saline (PBS), the cells were analyzed by flow cytometric analysis using a BD FACSAria™ II (BD Biosciences, San Jose, CA, USA). The data were analyzed with FlowJo software (version 7.6.2, by Flowjo LLC, OR, USA). We analyzed at least 50,000 events per sample and calculated the numbers of different subsets of circulating memory Tfh cells in individual samples according to the counts of lymphocytes per liter of blood multiplied by the percentage of different subsets of memory Tfh cells in lymphocytes.
Pe cy 7 anti ccr7
Manufactured by BD
Sourced in United States
PE)-Cy™7-anti-CCR7 is a fluorochrome-conjugated antibody that binds to the CCR7 receptor. It is used in flow cytometry applications to identify and analyze cells expressing the CCR7 protein.
Automatically generated - may contain errors
Lab products found in correlation
Percp cy5.5 anti cxcr5, by BD (2 mentions)
Anti icos pe, by BD (2 mentions)
Anti pd 1 bv421, by BD (2 mentions)
Pe cf594 anti cd154, by BD (2 mentions)
Facsaria 2, by BD (2 mentions)
Anti cd4 bv510, by BD (1 mentions)
Alexa fluor 488 anti samhd1, by Euromedex (1 mentions)
Rabbit anti samhd1, by Euromedex (1 mentions)
Alexa fluor 647 antirabbit antibody, by Thermo Fisher Scientific (1 mentions)
Pe anti bcl 6, by BD (1 mentions)
Pe ki67, by BD (1 mentions)
Anti foxp3 pe, by BioLegend (1 mentions)
Pe cf594 anti cd45ro, by BD (1 mentions)
Alexa fluor 647 anti ccr4, by BD (1 mentions)
Anti cxcr3 pe, by BD (1 mentions)
Pe cy7 anti ccr6, by BD (1 mentions)
Foxp3 permeabilization solution kit, by Thermo Fisher Scientific (1 mentions)
Live death vivid detection kit, by Thermo Fisher Scientific (1 mentions)
Lsrii flow cytometer, by BD (1 mentions)
3 protocols using pe cy 7 anti ccr7
1
Quantification of Circulating Memory Tfh Cells
2
Characterization of Memory T Follicular Helper Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human PBMCs at 106/tube were stained in duplicate with APC-H7-anti-CD3, BV510-anti-CD4, FITC-anti-CD45RA, PE-Cy7-anti-CCR7, PerCP-Cy5.5-anti-CXCR5, PE-anti-ICOS, BV421-anti-PD-1, PE-CF594-anti-CD154, or isotype-matched control IgG (Becton Dickinson, San Diego, USA) at room temperature for 30 minutes, respectively. After being washed with PBS, the cells were subjected to flow cytometry analysis in a BD FACSAria Ⅱ (BD Biosciences, San Jose, CA USA) and data were analyzed with FlowJo software (v7.6.2). At least 50,000 events per sample were analyzed. The cells were gated initially on living lymphocytes, on CD3+ and CD4+, and then on CXCR5+ and CD45RA-. Subsequently, the frequency of ICOS+, PD-1+ or CD40L+ CD3+CD4+CXCR5+CD45RA- T (memory Tfh) cells were analyzed. In addition, the CD3+CD4+CXCR5+CD45RA- Tfh cells were gated on CCR7 and the frequency of ICOS+, PD-1+, CD40L+ CCR7+ or CCR7- Tfh cells was analyzed. The numbers of each type of memory Tfh cells in individual subjects were calculated by the counts of lymphocytes per liter of blood multiplied by the percentage of different subsets of memory Tfh cells in lymphocytes.
3
Multicolor Flow Cytometry of T-cell Subsets
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Multicolor flow cytometry was performed on fresh PBMCs and frozen LNMCs. T-cell subpopulations were determined using the different combinations of the following fluorochrome-conjugated antibodies: Pacific Blue, PE-Cy7 or Alexa-Fluor 700anti-CD3, PerCP or Alexa-Fluor 700 anti-CD4, PE anti-CD27, PE-Cy7 anti-CCR7, PE-CF594 anti-CD45RO, Alexa-Fluor 488 anti-CXCR5, Alexa-Fluor 647 anti-CCR4, PE anti-CXCR3, PE-Cy7 anti-CCR6 (Becton Dickinson Biosciences, Pont de Claix (Le), France). Vioblue anti-CD3, APC-Vio770 anti-CD4, APC-Vio770 anti-CD45RO, APC anti-CD28 (Milteniy Biotech, Paris, France). Brillant Violet 421 anti-CD279 (Biolegend, London, UK).
Intracellular staining was carried out using the FoxP3 permeabilization solution kit according to the manufacturer's instructions (eBioscience, Paris, France) together with PE Ki67, PE anti-Bcl-6 (BD Pharmingen) or PE anti-FoxP3 (Biolegend), and Alexa-Fluor 488 anti-SAMHD1 (a gift from O. Schwartz) or rabbit anti-SAMHD1 (Euromedex, Souffelweyersheim, France) followed by Alexa-Fluor 647 antirabbit antibody (Invitrogen, Life Technologies, Saint Auben, France).
Dead cells were excluded using the Live/Death Vivid detection kit labeled with an aqua dye (Invitrogen). Fluorescence intensities were measured with an LSR II flow cytometer (Becton Dickinson) and analyzed using FlowJo version 7.6.5 (Tree Star Inc., Ashland, Oregon, USA).
Intracellular staining was carried out using the FoxP3 permeabilization solution kit according to the manufacturer's instructions (eBioscience, Paris, France) together with PE Ki67, PE anti-Bcl-6 (BD Pharmingen) or PE anti-FoxP3 (Biolegend), and Alexa-Fluor 488 anti-SAMHD1 (a gift from O. Schwartz) or rabbit anti-SAMHD1 (Euromedex, Souffelweyersheim, France) followed by Alexa-Fluor 647 antirabbit antibody (Invitrogen, Life Technologies, Saint Auben, France).
Dead cells were excluded using the Live/Death Vivid detection kit labeled with an aqua dye (Invitrogen). Fluorescence intensities were measured with an LSR II flow cytometer (Becton Dickinson) and analyzed using FlowJo version 7.6.5 (Tree Star Inc., Ashland, Oregon, USA).
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!