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 filter 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).
Beta plate scintillation mixture
Manufactured by PerkinElmer
Beta plate scintillation mixture is a specialized liquid scintillation cocktail used in beta particle detection and quantification. It is designed to be used in conjunction with beta plate counters or similar instrumentation to measure the radioactivity of samples. The mixture contains organic scintillators and other components that facilitate the detection of beta particles emitted from radioactive samples.
Automatically generated - may contain errors
Lab products found in correlation
1205 betaplate, by PerkinElmer (4 mentions)
3h thymidine, by MP Biomedicals (4 mentions)
Type a lter mats, by PerkinElmer (3 mentions)
Rpmi 1640 10 fetal bovine serum, by Thermo Fisher Scientific (3 mentions)
Type a filter mats, by PerkinElmer (2 mentions)
Fcm calibur ow cytometer, by BD (2 mentions)
Thymidine, by PerkinElmer (1 mentions)
96 well flat bottom culture plates, by Greiner (1 mentions)
5 protocols using beta plate scintillation mixture
1
Measuring Cell Proliferation in MLR
2
Leukocyte Activation and Proliferation Assay
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Leukocyte activation was performed using 96-well flat-bottom culture plates (Greiner Bio-One; Germany) for 72 h at 37 °C in the presence of 5% CO2. For standard two-way MLR assays, responder cells (total splenocytes, 2 × 105) were co-cultured with an equal number of stimulator splenocytes in 200 μl medium14 (link),16 (link). Thymidine (1 μ Ci/well; PerkinElmer Life and Analytical Sciences) was added 18–24 h before recovering (Inotech Biosystems International Inc.) using Type A filter mats (PerkinElmer Life and Analytical Sciences) and a beta-plate scintillation mixture (PerkinElmer). CPM were determined using a liquid scintillation analyzer (Packard 1900CA, Packard Instrument Co.). Data were expressed as the mean CPM of triplicate determination, and converted into proliferation percentages. 100% proliferation refers to vehicle treated allogeneic group. Splenocytes background readout values (medium alone) were deducted from the results.
3
Characterizing T Cell Activation in Mixed Lymphocyte Reaction
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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.
[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.
4
Comprehensive Immune Response Profiling
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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.
[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.
5
Mixed Lymphocyte Reaction and Cytokine Quantification
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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.
[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.
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