WST-1 Cell Proliferation and Cytotoxicity Assay Kit

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Description

Catalog No.: CSK1158
Introduction
1. WST-1 Cell Proliferation and Cytotoxicity Assay Kit is a sensitive and accurate assay for cell
cytotoxicity and proliferation. The kit components are sufficient for performing up to 100 assays.
2. WST-1 assay is much like MTT assay and the MTS assay, they are colorimetric assays for
measuring the activity of enzymes that reduce MTT or close dyes (XTT, MTS, WSTs) to formazan
dyes, giving a purple yellow. A main application allows assessing the viability (cell counting) and the
proliferation of cells (cell culture assays). It can also be used to determine cytotoxicity of potential
medicinal agents and toxic materials, since those agents would stimulate or inhibit cell viability
and growth.
3. This kit is very convenience to use. It is unnecessary to use isotope, to wash and collect cells, as
well as to solute formazan. The entire step can be done in only one 96-well plate.
4. No interrupt with Phenol red and serum.
5. WST-1 is not toxic to cells. After adds WST-1, it can be read by coated wells repeatedly in different
time. In this way, the detect time will be more flexible to find the best detect time.
6. The kit components are sufficient for performing up to 100 assays.
Kit Component
WST-1?powder? 1 tube
Electron coupling reagent 1ml
Storage
At -20? in dark for one year. After reconstituting, WST-1 solution can be stored at 4? in dark for one
week, at -20? in dark for half a year?avoid repeatedly freezing and thawing?.
Note
1. If cell culture time is too long, please pay attention to the evaporation issue. There are two solutions
for this. a. Avoid using the outmost wells of the plate, and add PBS buffer, water or culture fluid
instead; b. or place 96 wells near by the water in incubator.
2. Please wear transparent gloves when operate.

Protocol
1. WST-1 preparation: add 1ml Electron coupling reagent into WST-1 powder and mix thoroughly.
After dissolving, the frozen WST-1 solution may appear some sediments, so, try to incubate it
for 2-10 min at 37? before use.
2. Collect logarithmic phase cell, adjust concentration of cell suspension; add cell suspension into
the plate, 100ul per well. In general, cells seeded at densities between 1000-10,000 cells per
well (side holes filled with aseptic PBS buffer). Seed cells in a 5% Co2 incubator at 37? until
cells bespread well bottom for one floor (The volume of cell for each well should be determined
according to cells’ size and breed speed). Add concentration gradient drug. Principlely, add
drug after cells adhere. 0-10 ?l per well. Using 3-5 repeating pipettors.
3. Add 10 ?l WST-1 Reagent to each well, continue to culture for 4 hours. If drug react with
WST-1, you could centrifugal first then remove nutrient solution. Wash with PBS buffer carefully
2-3 times; add nutrient solution with WST-1.
4. The incubate time response to the situation of cell’s type and concentration. The first
experiment can try to read the result after 0.5 hour, 1hour, 2 hours and 4 hours solely., then
chose a proper time for next step.
5. Shake 96 wells for 1 min until solution become homogeneous.
6. Read absorbance at 450nm. If there’s no 450nm filter, use 420-480nm instead. During dual
wavelength spectrophotometry, you may choose wavelength longer than 600nm.