In recent years, cell proliferation has been a hot topic in the field of biomedicine. Cell proliferation refers to the series of complex reactions such as DNA replication, RNA transcription, and protein synthesis that occur under the regulation of cyclic control factors, with the replication and doubling of nuclear DNA being a significant characteristic of the entire process.

The detection of cell proliferation is widely used in research fields such as molecular biology, tumor biology, pharmacology, and pharmacokinetics. It is not only very important in studying the basic biological characteristics of cells but also a fundamental method for analyzing cell status and researching genetic traits, providing valuable information for exploring the pathogenesis of diseases, diagnosing diseases, and treating diseases.

The methods for detecting cell proliferation are currently mainly divided into metabolic activity detection, DNA synthesis detection, and detection of cell proliferation-related antigens. In addition, enzyme activities such as ATP, calcium ion influx, and cell membrane integrity can also be detected. Each detection method has its own advantages and disadvantages. Different experimental purposes should be considered to choose detection methods with high accuracy and good repeatability, or to combine multiple detection methods to objectively and comprehensively reflect the results of the experiment.Xiao Ai mainly introduces two commonly used methods for detecting cell proliferation: metabolic activity detection and DNA synthesis detection.

I. Metabolic Activity Detection

✦ CCK-8 Kit abs50003

Product Introduction: The Cell Counting Kit 8, referred to as CCK-8, is a highly sensitive, non-radioactive colorimetric method for determining the number of viable cells in proliferation or cytotoxicity assays.

Principle of Detection: CCK-8 is based on the water-soluble tetrazolium salt WST-8. In the presence of an electron-coupling reagent, WST-8 can be reduced by dehydrogenases in the mitochondria to form an orange-yellow formazan dye. The formazan dye is soluble in tissue culture medium and is proportional to the number of viable cells. By colorimetry, the number of viable cells can be dynamically quantified, allowing for the detection of cell proliferation or drug toxicity. This kit is typically used for the detection of cell cycle and apoptosis in adherent or suspension cells. If used for tissues, the tissue must be digested into a single-cell state before testing.

Product Advantages: The CCK-8 solution can be directly added to cell samples without the need to prepare various components in advance. It is fast, highly toxic, and cost-effective. It is widely used in drug screening, cell proliferation assays, cytotoxicity assays, tumor drug sensitivity tests, and the detection of biofactor activity.

Cited Literature: Absin CCK-8 Kit has been cited in a total of 59 publications, including "Biomimetic Immunosuppressive Exosomes that Inhibit Cytokine Storms Contribute to the Alleviation of Sepsis" with an Impact Factor (IF) of up to 30.849.

 

✦ MTT Cell Proliferation and Cytotoxicity Detection Kit abs50010

Product Introduction: The MTT assay, also known as the MTT colorimetric assay, is a method for detecting cell viability and growth.

Principle of Detection: The succinate dehydrogenase in the mitochondria of living cells can reduce the exogenous MTT to an insoluble blue-purple formazan crystal that is deposited within the cells, a function that dead cells lack. The amount of MTT crystal formed is proportional to the number of cells within a certain range. The higher the absorbance, the stronger the cell activity, indicating lower drug toxicity.

Product Advantages: It is highly sensitive and cost-effective. It has been widely used in the detection of bioactive factors, large-scale antitumor drug screening, cytotoxicity tests, and the determination of tumor radio-sensitivity.

Cited Literature: "Long noncoding RNA MEG3 suppresses cell proliferation, migration and invasion, induces apoptosis and paclitaxel-resistance via miR-4513/PBLD axis in breast cancer cells", with an Impact Factor (IF) of 3.699.

✦ Alamar Blue Cell Proliferation and Toxicity Detection Reagent abs47047610

Product Introduction: Alamar Blue is a cell viability assay reagent that contains resazurin, a non-toxic and weakly blue-fluorescent indicator that is cell membrane permeable.

Principle of Detection: Resazurin is a redox indicator that changes color based on the metabolic reduction by cells. The oxidized form of resazurin is blue-purple and essentially non-fluorescent, while its reduced product, resorufin, turns pink and highly fluorescent, with fluorescence intensity proportional to the number of respiratory active cells. The color change of Alamar Blue can be detected by a standard spectrophotometer by measuring absorbance changes at 570nm with a reference wavelength of 600nm; the fluorescence changes can be detected by a fluorescence spectrophotometer with excitation wavelengths between 530~560nm and an emission wavelength of 590nm.

Product Advantages: Alamar Blue is a single reagent that allows for continuous and rapid detection of cell proliferation. It is non-toxic and non-interfering with the electron transport chain, cell respiration or function, and does not affect antibody synthesis and secretion. It is suitable for continuous observation of cell proliferation in the same batch of cells and for further observation, with simple operation and minimal interference with normal cell metabolism.

II. Frequently Asked Questions (FAQ)

Question	Answer
Q1: How long should adherent cells be pre-cultured when using the CCK-8 kit?	A1: It depends on the type of cells. For the first use, it is necessary to explore the conditions. For adherent cells, the culture time with CCK-8 is generally 1-4 hours, but the color development can be visually observed after about 30 minutes of incubation.
Q2: How many cells should be seeded for the CCK-8 kit to detect cell proliferation?	A2: It varies with different types of cells. For the first experiment, it is recommended to start with a few wells to determine the number of cells to seed and the incubation time after adding CCK-8 reagent.
Q3: How can the experiment ensure the same number of cells seeded per well?	A3: When seeding, ensure that the cell suspension is well mixed to avoid cell sedimentation, which could lead to unequal cell numbers per well. Mix the suspension every few wells.
Q4: How to determine cell toxicity or proliferation based on the OD value?	A4: Assays like MTT or CCK-8 reflect toxicity by measuring viable cells. A decrease in OD value compared to the control group indicates lower cell viability, higher toxicity, or inhibited proliferation.
Q5: My cell culture medium contains phenol red. Will it affect the experimental results?	A5: The presence of phenol red in the culture medium does not affect the CCK-8 kit's measurement of cell viability.
Q6: Can the CCK-8 kit be used for检测 ("detection" may be more appropriate here, as "检测" can also mean "inspection" or "examination") of cells in highly turbid culture medium (检测 "detection" of cells in highly turbid culture medium)? Will it affect the experimental results? (Will it affect the experimental results?)	A6: If the sample is a highly turbid cell suspension, it is recommended to set 600 nm (or above) as the reference wavelength and subtract the OD value at the reference wavelength.

 

Product Number	Product Name	Specification	Lead Time
abs47047610	Alamar Blue Cell Proliferation and Toxicity Detection Reagent	500T/1000T/5000T	1-2 weeks
abs50003	CCK-8 Kit	500T/5000T	In Stock
abs50010	MTT Cell Proliferation and Cytotoxicity Detection Kit	500T	In Stock

Note: All Absin products are for research use only and should not be used for medicinal, household, or other purposes.

 

 

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