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Catalog No. CSK1175Size 100 assaysKit ComponentTrypan Blue Staining solution (2X) 10ml(Dilution: 1:2)Cell diluents buffer 100mlStorageStore at 4℃ for one year.IntroductionTrypan blue, as one of cell staining dyes, hasbeen proved to be excluded by alive cells, whilecould stain dead cells to blue. Through an opticalmicroscope, the staining process can beobserved and the number of cells can bequantitative counted. But whether the cells aresuicide or are destroyed, this cannot be judged bythis stain.Alphabioregen’s Trypan Blue Staining CellViability Assay Kit is developed on this workingprinciple. Usually, the cell whose cytomembranelost its integrity is considered as a dead cell.After staining with trypan blue, users can recordthe number of cells directly through microscope,or take a photo under microscope then record thenumber. In this way, the accurate cell livability canbe quantified.
Protocol:1. Cell CollectionA. Digest anchorage-dependent cell with pancreatin or EDTA;B. For suspending cells, collect directly. And centrifuge the collected cells at 1000-2000g for 1 min, thenabandon the supernatant;C. Collect cell suspension solution, dilute it properly with cell diluents buffer (kit component); orcentrifuge and abandon the supernatant, estimate the number of cells. After that, add some cell diluentsbuffer (kit component) into the sediment and blew the re-suspending cells.2. StainingPipette 100μl cell suspension solution into EP tube, add 100μl trypan blue staining working solution (Kitcomponent and should be diluted 1:2 before use), then blew slightly and mix them thoroughly. 3-5 minlater (staining time should not be too long), drip this solution on the cell counting plate to calculate thenumber.3. Cell Viability AnalysisPipette the cell suspension solution which was added the trypan blue staining working solution (Kitcomponent and should be diluted 1:2 before use) onto the blood cell counting plate to count the number.Calculate the total number of cells and the number of cells dyed to blue respectively in the big quadrel.Cell Viability = (total number of cells- blue cell number)/ total number of cells X 100%Note1. This kit can be enough for 100 assays.2. The concentration of cell sample should better be 1X106/ml.3. Control the staining time well, otherwise cells would be poisoned to die.4. This product can be used to stain the growing anchorage-dependent cell.5. When calculate cell number, the cell dilution 104should be multiplied.Hemocymeter calculation method:Cell number of 1 mm quadrel X 104(cell dilution) = actual cell number/ml