Whole blood was collected in four 10ml EDTA collection tubes from each subject. PBMCs were isolated using Ficoll-Paque density gradient and stained with conjugated monoclonal antibodies against CD45 FITC, CD19 PE, CD45RA PE-CY7 (all BD Pharmingen), CD3 Brilliant Violet 421, CD4 CF594 (both BD Horizon), CD14 APC (BD Biosciences) and CD27 APC-eFlour780 (ebioscience). Cells were then stored overnight in buffer at 4° C and sorted the following day, on a BD FACSAria cell sorter (BD Biosciences). The following populations were gated for sorting following exclusion of debris and doublets: monocytes (CD45+CD14+); B cells (CD45+CD14-CD3-CD19+); naïve CD4+ T cells (CD45+CD14-CD19-CD3+CD4+CD27+CD45RA+) and memory CD4+T cells (CD45+CD14-CD19-CD3+CD4+CD45RA-). Cell counts and purity checks were performed after sorting, and then cells were stored frozen as a pellet at −80°C.
Cd4 cf594
Manufactured by BD
The CD4 CF594 is a fluorescent-labeled antibody reagent used for the identification and enumeration of CD4-positive T cells in flow cytometry applications. The CF594 fluorescent label is excited by the red laser and emits light in the far-red region of the spectrum.
Automatically generated - may contain errors
2 protocols using cd4 cf594
1
Isolation and Sorting of PBMC Subsets
2
Immune Cell Isolation and Sorting
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Whole blood was collected from each subject into four 10‐ml EDTA collection tubes. PBMCs were isolated using a Ficoll‐Paque density gradient and stained with conjugated monoclonal antibodies against CD45 (fluorescein isothiocyanate conjugated), CD19 (phycoerythrin [PE] conjugated), CD45RA (PE–Cy7 conjugated) (all from BD PharMingen), CD3 (Brilliant Violet 421 conjugated), CD4 (CF594 conjugated) (both from BD Horizon), CD14 (allophycocyanin [APC] conjugated) (BD Biosciences), and CD27 (APC–eFlour 780 conjugated) (ebioscience). Cells were then stored overnight in buffer at 4°C and sorted the following day, on a BD Biosciences FACSAria fluorescence‐activated cell sorter (FACS). The following populations were gated for sorting following exclusion of debris and doublets: monocytes (CD45+CD14+), B cells (CD45+CD14 −CD3 −CD19+), naive CD4+ T cells (CD45+CD14 −CD19 −CD3+CD4+CD27+CD45RA+), and memory CD4+ T cells (CD45+CD14 −CD19 −CD3+CD4+CD45RA−). Cell counts and purity checks were performed after sorting, and then cells were stored frozen as a pellet at −80°C.
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!