Fcm calibur ow cytometer

Cells activated in an MLR were allowed to incubate for up to 3 days before harvesting.
[3H]Thymidine(0.5μCi/well; MPBiomedicals) was added 24h before harvesting (SkatronInstruments) using Type A lter mats (Perkin-Elmer Life and Analytical Sciences) and a beta plate scintillation mixture (Perkin-Elmer). Disintegrations per minute were determined using a liquid scintillation counter (1205 Betaplate; Perkin-Elmer).
ELISA assay for quantitating cytokine production X-ray-irradiated DC (2×10 4 ) and the T cells (1×10 5 ) were seeded into wells of 96-well at-bottom culture plates in RPMI 1640 10% fetal bovine serum (Invitrogen Life Technologies) for 3 days. Supernatants were removed and further analyzed for cytokine production (IL-2 and IFN-γ) with ELISA. Plates were read at 405 nm using a Labsystems Multiskan enzyme-linked immunosorbent assay reader.
Flow cytometry for detecting activation marker expression FCM assay was used to characterize activation marker expression of T cells. CD4 + or CD8 + T cells were cocultured with DCs for 3 days. The cells were washed with PBS and then stained with anti-CD3-PE, anti-CD25-FITC and anti-CD69-APC at 4 •C for 1h, and then washed by PBS. CD25 and CD69 activation markers were measured using a FCM Calibur ow cytometer (BD company). Data were analyzed with CellQuest software.

Cells activated in an MLR were allowed to incubate for up to 3 days before harvesting.
[3H]Thymidine(0.5 µCi/well; MPBiomedicals) was added 24 h before harvesting (SkatronInstruments) using Type A lter mats (Perkin-Elmer Life and Analytical Sciences) and a beta plate scintillation mixture (Perkin-Elmer). Disintegrations per minute were determined using a liquid scintillation counter (1205 Betaplate; Perkin-Elmer).
ELISA assay for quantitating cytokine production X-ray-irradiated DC (2 × 10 4 ) and the T cells (1 × 10 5 ) were seeded into wells of 96-well at-bottom culture plates in RPMI 1640 10% fetal bovine serum (Invitrogen Life Technologies) for 3 days. Supernatants were removed and further analyzed for cytokine production (IL-2 and IFN-γ) with ELISA. Plates were read at 405 nm using a Labsystems Multiskan enzyme-linked immunosorbent assay reader.
Flow cytometry for detecting activation marker expression FCM assay was used to characterize activation marker expression of T cells. CD4 + or CD8 + T cells were cocultured with DCs for 3 days. The cells were washed with PBS and then stained with anti-CD3-PE, anti-CD25-FITC and anti-CD69-APC at 4 •C for 1 h, and then washed by PBS. CD25 and CD69 activation markers were measured using a FCM Calibur ow cytometer (BD company). Data were analyzed with CellQuest software.

Loading [MathJax]/jax/output/CommonHTML/fonts/TeX/fontdata.js FCM assay was used to characterize activation marker expression of T cells. CD4 + or CD8 + T cells were cocultured with DCs for 3 days. The cells were washed with PBS and then stained with anti-CD3-PE, anti-CD25-FITC and anti-CD69-APC at 4 •C for 1h, and then washed by PBS. CD25 and CD69 activation markers were measured using a FCM Calibur ow cytometer (BD company). Data were analyzed with CellQuest software.