Peripheral blood mononuclear cells (2 × 106/ml) were incubated with 0 or 1 μg/ml plate-bound purified mouse anti-human CD3 antibody (Life Technologies) in flat-bottomed 96-well plates. After 54 h in culture, the cells were pulsed with [3H] thymidine (1 μCi/well; Perkin-Elmer, Waltham, MA, USA) and harvested 18 h later. Following incubation, cells were harvested onto glass fiber filter mats (Perkin-Elmer) via a MicroBeta FilterMate-96 Harvester (Perkin-Elmer). Incorporated radioactivity was measured by liquid scintillation counting on a 2450 MicroBeta plate counter (Perkin-Elmer). Each assay was performed in triplicate. The proliferative response was expressed as a stimulation index (SI) calculated by dividing the mean counts per minute (cpm) of anti-CD3-stimulated T cells by the mean cpm of unstimulated (media only) cells.
Glass fiber filter mats
Manufactured by PerkinElmer
Sourced in United States
Glass fiber filter mats are a type of laboratory equipment used for filtration purposes. They are composed of fine glass fibers that are woven or bonded together to create a porous material. The primary function of these filter mats is to separate solid particles from liquid or gaseous samples, allowing the filtrate to pass through while retaining the particulates.
Automatically generated - may contain errors
Lab products found in correlation
3h thymidine, by PerkinElmer (2 mentions)
Microbeta filtermate 96 harvester, by PerkinElmer (1 mentions)
Mouse anti human cd3 antibody, by Thermo Fisher Scientific (1 mentions)
β scintillation counter, by PerkinElmer (1 mentions)
Wallac microbeta trilux, by PerkinElmer (1 mentions)
Celltiter 96 aqueous non radioactive cell proliferation assay kit, by Promega (1 mentions)
Mrx 2, by Dynex (1 mentions)
3h tdr, by PerkinElmer (1 mentions)
4 protocols using glass fiber filter mats
1
T Cell Proliferation Assay
2
B Cell Proliferation Assay
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Negatively selected purified B cells were cultured in appropriate conditions in 96-well U-bottom plates (Costar) for 24h. 3H-thymidine (0.5 μCi/ well; Perkin Elmer) was added to the cultures and incubated for a further 16h. The cells were then harvested onto glass fiber filter mats (Perkin Elmer) using a Tomtec Harvester. Proliferation was measured as 3H- thymidine incorporation using a Perkin Elmer β-scintillation counter. Results were expressed as counts per minute (CPM).
3
Evaluation of Tumor Cell Viability and Proliferation
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Cell viability was assessed by MTS assay CellTiter 96® AQueousNon-Radioactive Cell Proliferation Assay kit (Promega, Madison, WI, USA), while
cell proliferation was measured using tritiated thymidine (3H-TdR)
incorporation assay. Various ratio of tumor cells with UC-MSCs (1:0.2, 1:0.1,
1:0.02, and 1:0.01) were co-cultured with a fixed number of leukemic tumor cell
lines (0.5 × 105 cells/well) in 96-well plates in complete RPMI 1640
medium for 3 days. For the viability assay, 30 µl of MTS mixed with PMS
(phenazine methosulfate) was added to each well and incubated for 3–4 h. The
absorbance values of tumor cells alone and tumor cells in co-culture with
UC-MSCs were measured by using a microtiter plate reader (Dynex Technologies,
MRX II, Chantilly, VA, USA) at 490 nm. For the tritiated thymidine
(3H-TdR) assay, 3H-TdR (0.037 MBq/well [0.5 µCi/well])
(Perkin Elmer, Waltham, MA, USA) was pulsed to the cells at the final 18 h of
incubation. At 72 h, cells were harvested using an automated cell harvester
(Harvester 96 MACH III M, TOMTEC, Hamden, CT, USA) onto glass fiber filter mats
(Perkin Elmer, Waltham, MA, USA). After drying the filter mats, scintillation
fluid was added, and the DNA content was measured by thymidine incorporation
using the beta counter (WALLAC Micro Beta Trilux, Perkin Elmer, Waltham, MA,
USA).
cell proliferation was measured using tritiated thymidine (3H-TdR)
incorporation assay. Various ratio of tumor cells with UC-MSCs (1:0.2, 1:0.1,
1:0.02, and 1:0.01) were co-cultured with a fixed number of leukemic tumor cell
lines (0.5 × 105 cells/well) in 96-well plates in complete RPMI 1640
medium for 3 days. For the viability assay, 30 µl of MTS mixed with PMS
(phenazine methosulfate) was added to each well and incubated for 3–4 h. The
absorbance values of tumor cells alone and tumor cells in co-culture with
UC-MSCs were measured by using a microtiter plate reader (Dynex Technologies,
MRX II, Chantilly, VA, USA) at 490 nm. For the tritiated thymidine
(3H-TdR) assay, 3H-TdR (0.037 MBq/well [0.5 µCi/well])
(Perkin Elmer, Waltham, MA, USA) was pulsed to the cells at the final 18 h of
incubation. At 72 h, cells were harvested using an automated cell harvester
(Harvester 96 MACH III M, TOMTEC, Hamden, CT, USA) onto glass fiber filter mats
(Perkin Elmer, Waltham, MA, USA). After drying the filter mats, scintillation
fluid was added, and the DNA content was measured by thymidine incorporation
using the beta counter (WALLAC Micro Beta Trilux, Perkin Elmer, Waltham, MA,
USA).
4
Cell Proliferation Assay with Tritiated Thymidine
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One hundred microliters of cultured cells were transferred to 96‐well U‐bottomed plates and pulsed with 0.5 μCi/well of 3H‐thymidine and incubated for 24 hours at 37°C, 5% CO2. The plates were harvested using a Tomtec harvester and the glass fiber filter mats (PerkinElmer, MA) were left to dry for at least 24 hours. The filter mats were treated with liquid scintillation fluid (Scintillant‐Beta plate scint; Wallac) and then counted using a Wallac scintillation counter to measure proliferation.
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