Peripheral blood mononuclear cells (PBMCs) were isolated from fresh blood samples obtained from BD patients and healthy controls using density gradient centrifugation (Amersham Biosciences, Uppsala, Sweden). Monocytes and CD4+ T cells were purified using magnetic bead separation from PBMCs (Miltenyi Biotec, Cologne, Germany). The purity of isolated cell populations was confirmed by flow cytometry analysis using fluorochrome-conjugated antibodies against CD14 for monocytes and CD4 for T-helper lymphocytes and was over 90% for both monocytes and CD4+ T cells. Genomic DNA was obtained from isolated monocytes and CD4+ T cells using the Qiagen DNeasy Blood and Tissue Kit.
Pbmcs
Manufactured by Miltenyi Biotec
Sourced in Germany, United States
PBMCs are a type of blood cell that includes lymphocytes and monocytes. They are commonly used in various biomedical research applications.
Automatically generated - may contain errors
Lab products found in correlation
Glutamax supplement, by Thermo Fisher Scientific (2 mentions)
Dneasy blood and tissue kit, by Qiagen (1 mentions)
Interleukin 2 (il 2), by Thermo Fisher Scientific (1 mentions)
Texmacs, by Miltenyi Biotec (1 mentions)
Human t cell transact, by Miltenyi Biotec (1 mentions)
Human pan t isolation kit, by Miltenyi Biotec (1 mentions)
Phorbol 12 myristate 13 acetate pma, by Merck Group (1 mentions)
Carboxyfluorescein succinimidyl ester (cfse), by Thermo Fisher Scientific (1 mentions)
Ionomycin, by Merck Group (1 mentions)
Flow cytometry, by BD (1 mentions)
Carboxyfluorescein succinimidyl ester (cfse), by BD (1 mentions)
Monensin solution, by BioLegend (1 mentions)
Cd4 t cell positive selection kit, by STEMCELL (1 mentions)
Dq ova, by Thermo Fisher Scientific (1 mentions)
Dq ovalbumin, by Thermo Fisher Scientific (1 mentions)
6 protocols using pbmcs
1
PBMC Isolation and Cell Purification
2
Isolation and Activation of Human T Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
PBMCs (purchased from AllCells) were magnetically separated with Human Pan T Isolation Kit (Miltenyi Biotec). The newly isolated T cells were then cultured in complete T cell medium, composed of TexMACS (Miltenyi Biotec) supplemented with IL-2 (PeproTech, 100 IU/ml) and Penicillin Streptomycin (1:100). Cells were activated with human T cell TransACT (Miltenyi Biotec) for approximately 20 h before transfection.
3
BDCA1+ and BDCA3+ PBDC-Mediated T Cell Activation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Isolated BDCA1+ and BDCA3+ PBDCs were stimulated with 100 μg/mL CFP for 6 h. Syngeneic CD4 and CD8 T cells were isolated from PBMCs (Miltenyi Biotec) of the same donor and labeled with 10 μM carboxyfluorescein succinimidyl ester (CFSE) (Invitrogen, San Diego, CA, USA). The BDCA1+ PBDCs were co-cultured with CD4 T cells and BDCA3+ PBDCs were incubated with CD8 T cells a ratio of 1:25, respectively. Three days after co-culture, the cells were further stimulated for 4 h with 50 ng/mL phorbol 12-myristate 13-acetate (PMA; Merck, Kenilworth, NJ, USA) and 1 μM ionomycin (Merck), with the addition of monensin solution (BioLegend) during the final 2 h. The CFSE dilution and intracellular IFN-γ production in CD4+ and CD8+ T cells were analyzed using flow cytometry (Becton Dickinson).
4
Isolating HIV-Positive CD4 T Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Memory CD4 T cells from well-suppressed deidentified participants were isolated by negative selection from PBMCs (Miltenyi), incubated overnight in growth media (IMDM + 10% FBS) and stimulated overnight with 10 μg/mL Con A. The remaining CD4high T cells were then depleted using a CD4 T cell positive selection kit (StemCell). The unbound cells (enriched CD4−, HIV+ cells) were cultured overnight in growth media (IMDM + 10% FBS), concentrated by centrifugation, and seeded on poly-L-lysine coated coverslips and fixed using 4% formaldehyde for imaging.
5
Plasmacytoid Dendritic Cell Activation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
pDCs were isolated from PBMCs (Miltenyi Biotec) and were then cultured in RPMI medium 1640 with GlutaMAX supplement (Thermo Fisher Scientific) containing 10% (vol/vol) FBS and 100 U/ml penicillin/streptomycin in a 96-well plate. Purified pDCs were cultured with or without 10 μg/ml DQ OVA (Molecular Probes) in the presence or absence of 50 ng/ml TNF-α. After 18 h, the cells were collected and then washed twice before data were analyzed using flow cytometry.
6
Plasmacytoid Dendritic Cell Culture
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
pDCs were isolated from PBMCs (Miltenyi Biotec) and were then cultured in RPMI medium 1640 with GlutaMAX supplement (ThermoFisher Scientific) containing 10% (vol/vol) FBS and 100 U/mL penicillin/streptomycin in a 96-well plate. Purified pDCs were cultured with or without 10 μg/mL DQ Ovalbumin (Molecular Probes). After 18 h, the cells were collected and then washed twice before data were analyzed using flow cytometry. Samples from healthy controls (n = 3), At-Risk individuals (n = 3), and SLE patients (n = 3) were used.
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!