A proliferation of PBMCs was measured by XTT assay after 72 h of post-stimulation. The XTT working solution was prepared by adding 10 mg of XTT reagent (Biotium, USA) in 10 ml of RPMI media without phenol red (Sigma, USA) plus 100 μl of phenazine methosulfate (PMS, Sigma, USA). In culture plate, 50 μl of XTT solution was added in triplicate wells. Plates were shaken gently for mixing of dye and incubated at 37°C for 4 h. The absorbance was measured at 500 nm and at 600 nm for reference. Stimulation index (SI) was calculated as the ratio between optical density (OD) values of stimulated cells to unstimulated cells.
Xtt reagent
Manufactured by Biotium
Sourced in United States
The XTT reagent is a laboratory product that can be used for colorimetric analysis. It is a tetrazolium-based compound that undergoes reduction in the presence of metabolically active cells, resulting in the formation of a colored formazan product that can be measured spectrophotometrically.
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Lab products found in correlation
3 protocols using xtt reagent
1
Measuring PBMC Proliferation by XTT Assay
2
Transwell Assay for Tumor-Adipocyte Interactions
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Semipermeable transwell membrane inserts (0.4 µm, Corning Costar Inc., Tewksbury, MA, USA) containing 15,000 3T3L1 preadipocytes or mature adipocytes were placed above 2000 PanIN/PDAC cells in 24-well culture plates in 500 μl DMEM, 0.5%FCS with glucose/glutamine concentrations as indicated, cultured 72 h, transwells removed, and 50 μl XTT reagent (Biotium Inc., Hayward, CA, USA) added, cells incubated overnight, then 100 μl of media was removed and added to a 96-well assay plate (Corning Costar Inc., Tewksbury MA, USA, Cat#3795) and absorbance read at 450 nm with background subtraction reading at 650 nm on an Biotek Epoch plate reader (BioTek Inc., Winooski, VT, USA).
3
Transwell Co-culture Metabolic Assay
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