Cell Counting Kit-3D

1. Repeated freeze-thaw cycles of this kit may reduce assay performance. Although testing shows no significant impact after three cycles，for optimal results，aliquot the kit after the first thaw. Minor precipitation may occur during repeated thawing; allow the solution to equilibrate to room temperature and dissolve thoroughly. 2. Temperature affects signal intensity and stability. Both cells and CCK-3D solution must be equilibrated to room temperature before measurement. Thaw and mix the CCK-3D solution at room temperature (≤25°C) or in a water bath (20-25°C) prior to use. 3. When using 96-well plates for detection，evaporation must be carefully managed for prolonged cell culture. First，avoid using the outer ring of wells due to increased evaporation risk; instead，add an equivalent volume of PBS，water，or culture medium to these wells. Second，position the 96-well plate near a water source inside the incubator to mitigate evaporation. 4. 3D cells typically form spheroids within the 96-well plate. The orange-yellow formazan formed after adding CCK-3D solution aggregates on and around the cell spheroids. Before detection，gently tap the 96-well plate to distribute the formazan evenly throughout the cell culture medium. Failure to do so may result in biased signal values，affecting the accuracy of the detection results. 5. This assay relies on a dehydrogenase-catalyzed reaction. Therefore，reducing agents (e. g. ，certain antioxidants) may interfere with detection. If significant reducing agents are present in the test system，measures should be taken to remove them. 6. Only standard transparent 96-well or 384-well plates should be used for detection. Alternatively，cells cultured in 3D using ultra-low-binding dishes or plates designed for 3D culture can be transferred to standard transparent multiwell plates for detection. 7. Before reading with an ELISA reader，ensure no bubbles are present in any well，as they will interfere with the assay. Product Description: Cell Counting Kit-3D，abbreviated as CCK-3D kit or CCK3D kit，is a rapid and highly sensitive kit based on WST series chromogenic substrates for the detection of 3D cell viability，proliferation，and cytotoxicity. This kit is suitable for measuring absorbance at approximately 450 nm，and the detection can usually be completed in about 1 hour. It can be used for detecting cell proliferation induced by cytokines，cell cytotoxicity induced by cytotoxic agents such as anti-cancer drugs，or cell growth inhibition induced by certain drugs. Product Features: 1. This kit is very convenient to use. The kit contains only one tube of pre-prepared CCK-3D solution，requiring no further preparation. No isotopes are required，and all detection steps are completed in the same 96-well plate. There is no need to wash or collect cells，nor is there a need for additional steps to dissolve formazan. It can be used for the detection of large numbers of samples. 2. Phenol red and serum do not significantly affect the measurement with this kit. 3. This product has no significant toxicity to cells. After adding the CCK-3D solution for color development，the plate can be repeatedly read with a microplate reader at different time points，making the detection time more flexible and allowing for the determination of the optimal measurement time. 4. This product is not only suitable for the detection of small numbers of samples but also ideal for high-throughput screening of large numbers of samples. It can be used with 3D cell spheroids in cell culture media prepared using various culture methods，including hanging drop culture plates，ultra-low attachment cell culture plates，Matrix-Gel? or Matrigel-coated plates，and agarose-coated plates.