Cell Titer Turbo 3D Luminescent Cell Viability Assay,Absin_specification/price/image

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Usage	Reagent Preparation: 1. Melting of reagents: Take out the reagents the day before the experiment and place them at 4 °C overnight to melt. The reagent can also be taken out on the day of the experiment and melted at room temperature, or melted in a 22 °C water bath, but it should be noted that the water temperature should not exceed 25 °C. 2. Equilibrate to room temperature: If the reagent melts at non-room temperature conditions, it can be placed in a 22 °C water bath before use to ensure that the reagent is balanced to room temperature before being used for testing. Note: Generally, it takes about 10 minutes for 5mL packaging; The 50 mL packaging takes about 20 minutes. 3. Mix well: Gently invert the solution 5 times before use to mix the solution evenly. Detection Procedure: 1. Use a 96-well plate suitable for chemiluminescence detection, and add microtissue containing cell culture medium to it. Note: (1) The prepared sample volume and microtissue characteristics (e.g., size, quantity, culture days, etc.) should be optimized for experimental conditions; (2) The total ATP content of the sample should be kept below 10μM (determine the upper limit concentration of the linear range); (3) The multi-well plate must be compatible with the photometer used. 2. Add test compounds to experimental wells and incubate according to your culture protocol. Note: Ensure that the volume of sample and test compound is low enough to allow the addition of equal volumes of reagent and then mixing without inter-well contamination. 3. Equilibrate the plate and its contents to room temperature (22-25 °C) for about 30 minutes. 4. Add 3D Reagent equal to the volume of cell culture medium present in each well to the detection well (e.g., for a 96-well plate, add 100 μl of 3D Reagent to 100 μl of cell-containing culture medium). 5. The contents were vigorously mixed for 5 minutes to induce cell lysis. Note: Mixing is important for efficient ATP extraction from 3D microtissues. 6. Allow the plate to incubate at room temperature for another 25 minutes to stabilize the luminescence signal. 7. Record luminescence. Note: The non-uniform luminescence signal within the plate may be caused by temperature gradients, non-uniform cell seeding, or edge effects in multi-well plates.
Description	3D cell cultures are more physiologically relevant and better representative of tissues in vivo, allowing more accurate predictions of efficacy or toxicity of drug treatments. 3D cell models are increasingly being used to understand disease mechanisms and discover drug therapies. Compared with 2D culture, 3D cell culture can predict the efficacy or toxicity of drug treatment more accurately. 3D Luminescent Cell Viability Assay Kit is a cell viability detection reagent based on luciferase system. It uses ATP-dependent luciferase-catalyzed luciferin luminescence reaction to measure intracellular ATP content through chemiluminescence signal to detect cell viability. The detection has wide linear range, high sensitivity and good stability. Due to the high-purity luciferin substrate, thermostable luciferase and optimized reaction reagents in this kit, this product has stronger cell lysis ability and can be used for cell viability analysis of 3D microtissue culture. This product is added to the cell culture to lyse the cell mass and release ATP, which can produce the reaction as shown in the figure and send out a stable light signal. The luminescence intensity is directly proportional to the amount of ATP, that is, the number of viable cells, within a certain range, so this product can quantitatively detect the number of viable cells. PRODUCT FEATURES The 3D Luminescent Cell Viability Assay has high detection sensitivity and wide linear range, and is compatible with small sample detection and high-throughput screening of large samples. 1. Convenient and fast: The detection reagents provided in the kit are ready-to-use, with stable readings and fast detection speed. It only takes about 10 minutes to complete the detection; 2. High sensitivity: It has a higher signal-to-noise ratio when applied to 3D microtissue detection; 3. Fast: data can be recorded within 30 minutes or less after adding reagents; 4. The luminescent signal is more stable: the half-life of the luminescent signal is ultra-stable, usually > 3 hours. 5. Stronger lysis ability: The optimized reaction reagent has stronger cell lysis ability and is suitable for cell viability analysis of 3D microtissue culture; 6. High throughput: It is compatible with the detection of a small amount of samples and the high throughput screening of a large number of samples.
General Notes	1. Temperature: The reagent contains luciferase, and repeated freezing and thawing will affect its activity. It is recommended to store at-20 ℃ in the dark from light after dispensing. Reagents and cell samples should be balanced to room temperature before use to avoid the influence of enzyme catalytic effect; 2. Chemical factors: The reaction rate and luminous intensity of luciferase are affected by the chemical environment. There are differences in luminescence intensity and attenuation rate among different types of media and serum. In addition, high drug content may interfere with the luciferase reaction, thus affecting the luminescence signal. It is recommended to set up control wells of cell culture medium containing drugs to eliminate the interference of solvents; 3. Light sensitivity: This reagent is sensitive to light. If it is exposed to light during storage, it will accelerate the attenuation of luminous intensity. If the reagent is transferred from the original container, make sure to keep it protected from light; 4. ATP contamination: It is recommended to wear masks and latex gloves during operation to avoid contact with surfaces and equipment that may be contaminated. Avoid inserting the tip of the gun tip into the vial multiple times during operation.
Instructions	reagent preparation: 1, reagent melting: The day before the experiment, the reagent was removed and placed at 4 ° C overnight to melt. It is also possible to remove the reagent on the day of the experiment and melt it at room temperature or place it in a 22℃ water bath, but it should be noted that the water temperature should not exceed 25℃. 2, balance to room temperature: if the reagent melts under non-room temperature conditions, it can be placed in a 22 ° C water bath before use, to ensure that the reagent is balanced to room temperature before detection. Note: Generally, it takes about 10min for 5mL packaging; 50mL packaging takes about 20min. 3, mixing: Before use, gently invert 5 times to mix the solution evenly. detection steps: 1, use a 96-well plate suitable for chemiluminescence detection, in which microtissue containing cell culture medium is added. Note: (1) The preparation sample volume and microtissue characteristics (e.g., size, number, incubation days, etc.) should be optimized for the experimental conditions; (2) The total ATP content of the sample should be maintained at 10μ 2, Add the test compound to the experimental well and incubate according to your culture protocol. Note: Make sure the volume of the sample and test compound is low enough to allow the addition of an equal volume of reagent, followed by mixing, without interwell contamination. 3, Equilibrate the plate and its contents to room temperature (22-25 ° C) for about 30 minutes. 4, Add Cell Titer Turbo 3D Reagent equal to the volume of cell medium present in each well to the assay Wells (for example, for a 96-well plate, add 100μ l Cell Titer Turbo 3D Reagent was added to 100μ l in the medium containing the cells). 5, mix the contents vigorously for 5 min to induce cell lysis. Note: Mixing is important for efficient ATP extraction from 3D microtissues. 6, Allow the plate to incubate at room temperature for an additional 25 minutes to stabilize the luminescence signal. 7, record luminescence. Note: The non-uniform luminescence signal in the plate may be caused by temperature gradients, uneven cell seeding, or edge effects in the multi-well plate.
Storage Temp.	1. Store below-10 °C and protect from light. In order to ensure the best performance, it is recommended to place it at-70 ℃ for long-term storage. 2. This product can still remain stable after 6 cycles of repeated freezing and thawing. 3. Sub-packaging may lead to the risk of ATP contamination, so avoid sub-packaging.
Applications	It is suitable for activity detection of bioactive factors, large-scale anti-tumor drug screening, cell proliferation, cytotoxicity test, etc.

AntBio is a biotechnology group company dedicated to serving life sciences, aiming to help scientists accelerate research and improve work efficiency. AntBio provides comprehensive and high-quality reagent tools for basic research, drug development, and diagnosis, including research grade antibodies, proteins, biochemical reagents, and assay kits. These research tools are widely used in different segments of life science research. The group company currently consists of three brands, Absin, Starter-Bio and UA-Bio.