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Home > Cell Analysis > Cell Proliferation / Cytotoxicity >
*Estimated. Exact shipping date will be notified.
For Research Use Only Products
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Colorimetric microplate assay
One Solution type (ready-to-use solution)
No washing required
No radioisotopes or organice solvents required
Highly Sensitive |
Kit Contents
| Cell Counting Kit-SK, 1,000 Tests |
:5 ml bottle x 2 |
| Cell Counting Kit-SK, 3,000 Tests |
:5 ml bottle x 6 |
Storage Condition : 0-5ºC, protect from light
Shipping Condition : ambient temperature
Required Equipment and Materials
plate reader with 450 nm filter, 96-well culture plate; 10µl, 100-200 µl, and multi-channel pipettes; CO²
incubator
Product Description
Cell Counting Kit-SK (CCK-SK) is an alternative kit to CCK-8 for cells or cell lines with a low metabolism. The O.D. of CCK-8 for leukocyte cells is sometimes very low such as, 0.2-0.4 after 2-4 hours of incubation because of lower metabolism compared with adherent type cells. CCK-SK enables a shorter incubation time to obtain a higher O.D. increase within 30 minutes to 1 hour. Assay mechanisms and assay protocols are the same as CCK-8. Fig. 1 shows the sensitivity of various assays.
Fig. 2 Absorption spectrum of WST-8 formazan dye
Sensitivity Data of CCK-SK in Jurkat Cell
Fig. 3 The Sensitivity of Commercial Products
WST-8 is the highest sensitive dye for the cell based assay.
Fig. 4 Comparison of the sensitivity using tetrazolium dye
Example Data
Fig. 5 Data for Cell Proliferation and Cytotoxicity Assay
What is the difference between CCK-SK and CCK-8?
Since the sensitivity using CCK-SK is higher than using CCK-8, CCK-SK enables a shorter incubation for coloring reaction. CCK-SK is suitable for cell lines that require higher sensitivity such as leucocyte or leukemia cell lines.
How many cells should there be in a well?
At least 1,000 cells are necessary per well (100 μl medium). The recommended maximum number of cells per well for the 96-well plate is 25,000. If a 24-well or 6-well plate is used for this assay, please calculate the number of cells per well accordingly, and adjust the volume of the CCK-SK solution in a well to 10% of the total volume.
Does phenol red affect the assay?
No. The absorption value of phenol red in a culture medium can be removed by subtracting the absorption value of a blank solution from the absorption value of each well. Therefore, a medium containing phenol red is usable.
I do not have a 450 nm filter. What other filters can I use?
You can use filters with an absorbance between 430 nm and 490 nm, even though a 450 nm filter gives the best sensitivity.
Can this kit be used with a 384-well plate?
Yes, you can use this kit for a 384-well plate. Add CCK-SK solution with 1/10 volume of the culture medium in a well. If the volume of CCK-SK to be added is too small, dilute CCK-SK in half with the medium and add 1/5 of the volume of culture medium in a well.
Can this kit be used with a 24-well plate?
Yes. Add CCK-SK solution with 1/10 volume of the culture medium in a well.
Does CCK-SK stain viable cells?
No, it does not stain viable cells because a water-soluble tetrazolium salt (WST-8) is used for the CCK-SK solution. The electron mediator, 1-Methoxy PMS, receives electrons from viable cells and transfers the electrons to WST-SK in the culture medium. Since its formazan dye is also highly water soluble, CCK-SK cannot be utilized for cell staining.
How stable is CCK-SK?
CCK-SK is stable over 6 months at room temperature and one year at 0-5ºC with protection from light. For longer storage, we recommend storing the solution at -20ºC ; CCK-SK is stable over 2 years at -20ºC.
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All Products are applicable only for life science research. Not for diagnostic research use.
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