Trypan Blue Staining Cell Viability Assay Kit

Description

Catalog No. CSK1175
Size 100 assays
Kit Component
Trypan Blue Staining solution (2X) 10ml
(Dilution: 1:2)
Cell diluents buffer 100ml
Storage
Store at 4℃ for one year.
Introduction
Trypan blue, as one of cell staining dyes, has
been proved to be excluded by alive cells, while
could stain dead cells to blue. Through an optical
microscope, the staining process can be
observed and the number of cells can be
quantitative counted. But whether the cells are
suicide or are destroyed, this cannot be judged by
this stain.
Alphabioregen’s Trypan Blue Staining Cell
Viability Assay Kit is developed on this working
principle. Usually, the cell whose cytomembrane
lost its integrity is considered as a dead cell.
After staining with trypan blue, users can record
the number of cells directly through microscope,
or take a photo under microscope then record the
number. In this way, the accurate cell livability can
be quantified.

Protocol:
1. Cell Collection
A. 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, then
abandon the supernatant;
C. Collect cell suspension solution, dilute it properly with cell diluents buffer (kit component); or
centrifuge and abandon the supernatant, estimate the number of cells. After that, add some cell diluents
buffer (kit component) into the sediment and blew the re-suspending cells.
2. Staining
Pipette 100μl cell suspension solution into EP tube, add 100μl trypan blue staining working solution (Kit
component and should be diluted 1:2 before use), then blew slightly and mix them thoroughly. 3-5 min
later (staining time should not be too long), drip this solution on the cell counting plate to calculate the
number.
3. Cell Viability Analysis
Pipette the cell suspension solution which was added the trypan blue staining working solution (Kit
component 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%
Note
1. This kit can be enough for 100 assays.
2. The concentration of cell sample should better be 1X10
6
/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 10
4
should be multiplied.
Hemocymeter calculation method:
Cell number of 1 mm quadrel X 10
4
(cell dilution) = actual cell number/ml