CytoSelect™ MTT Cell Proliferation Assay

Produktnummer ABIN2344923

Kurzübersicht für CytoSelect™ MTT Cell Proliferation Assay (ABIN2344923)

Reaktivität: Säugetier

Nachweismethode: Colorimetric

Applikation: Proliferation Assay (ProA)

Proben: Cell Samples

Produktdetails

Verwendungszweck: CytoSelect™ MTT Cell Proliferation Assay provides a colorimetric format for measuring and monitoring cell proliferation.

Bestandteile:

1. MTT Cell Proliferation Assay Reagent : One 10 mL bottle of MTT reagent.
2. Detergent Solution : One 100 mL bottle of Detergent Solution.

Benötigtes Material:

1. Cells for measuring proliferation
2. Cell culture medium
3. 24-well or 96-well clear cell culture plates.
4. Microtiter plate reader capable measuring absorbance at 540-570 nm.

Anwendungsinformationen

Applikationshinweise: Optimal working dilution should be determined by the investigator.

Kommentare:

• Colorimetric format for measuring and monitoring cell proliferation
• Kit contains sufficient reagents for the evaluation of 960 assays in 96-well plates or 192 assays in 24-well plates
• Cell proliferation reagent can be used to detect proliferation in bacteria, yeast, fungi, protozoa as well as cultured mammalian and piscine cells

Protokoll: The kit contains sufficient reagents for the evaluation of 960 assays in 96-well plates or 192 assays in 24-well plates. Cells can be plated and then treated with compounds or agents that affect proliferation. Cells are then detected with the proliferation reagent, which is converted in live cells from the yellow tetrazole MTT to the purple formazan form by a cellular reductase . An increase in cell proliferation is accompanied by an increased signal, while a decrease in cell proliferation (and signal) can indicate the toxic effects of compounds or suboptimal culture conditions. The assay principles are basic and can be applied to most eukaryotic cell lines, including adherent and non-adherent cells and certain tissues. This cell proliferation reagent can be used to detect proliferation in bacteria, yeast, fungi, protozoa as well as cultured mammalian and piscine cells. . Chemical Structures of Yellow MTT and Purple Formazan Product in Living Cells.

Testdurchführung:

1. Prepare a cell suspension containing 0.1-1.0 x 106 cells/mL in medium.
2. Add 100 μL per well to a 96-well cell culture plate or 500 μL per well to a 24-well cell culture plate with or without the compound to be tested. Culture the cells for 24-96 hours at 37 °C and 5 % CO2 in a humidified incubator.
3. Add 10 μL of the CytoSelect™ MTT Cell Proliferation Assay Reagent to each well if using a 96- well plate, or 50 μL to each well of a 24-well plate.
4. Incubate plate at 37 °C and 5 % CO2 for 3-4 hours until purple precipitate is visible (cellular precipitate can be more precisely visualized under a light microscope) 3
5. Add 100 μL of Detergent Solution per well of a 96-well plate, or 500 μL per well of a 24-well plate.
6. Incubate at room temperature for 2 hours to overnight protected from light. Note: Longer incubations with Detergent Solution in the wells may result in precipitate or turbidity that can increase background. If precipitate is observed, warm the plate at 37 °C for 10-20 minutes and agitate to dissolve the precipitate.
7. Read absorbance using 540-570 nm as the primary wavelength.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Lagerung: RT/-20 °C

Informationen zur Lagerung: The MTT Cell Proliferation Assay Reagent is a clear yellow ready-to-use solution, and it should be stored at -20°C protected from light. Store the Detergent Solution at room temperature. If precipitate or turbidity is observed in the Detergent Solution, warm the solution to 37°C for 10-20 minutes and agitate to dissolve the precipitate prior to use.

Referenzen

Guzelant, Isyar, Yilmaz, Sirin, Cakmak, Mahirogullari: "Are chondrocytes damaged when rheumatologic inflammation is suppressed?" in: Drug and chemical toxicology, Vol. 40, Issue 1, pp. 13-23, (2016) (PubMed).

Gumustas, Yilmaz, Isyar, Sirin, Batmaz, Ugras, Oznam, Ciftci, Mahirogullari: "Assessing the negative impact of phenyl alkanoic acid derivative, a frequently prescribed drug for the suppression of pain and inflammation, on the differentiation and proliferation of chondrocytes." in: Journal of orthopaedic surgery and research, Vol. 11, Issue 1, pp. 70, (2016) (PubMed).

Kim, Neznanov, Wilfong, Fleyshman, Purmal, Haderski, Stanhope-Baker, Burkhart, Gurova, Gudkov, Skitzki: "Preclinical Validation of a Single-Treatment Infusion Modality That Can Eradicate Extremity Melanomas." in: Cancer research, Vol. 76, Issue 22, pp. 6620-6630, (2016) (PubMed).

Isyar, Gumustas, Yilmaz, Sirin, Tosun, Mahirogullari: "Are We Economically Efficient Enough to Increase the Potential of in Vitro Proliferation of Osteoblasts by Means of Pharmacochemical Agents?" in: The open orthopaedics journal, Vol. 10, pp. 420-430, (2016) (PubMed).

Gumustas, Yilmaz, Sirin, Gumustas, Batmaz, Isyar, Akkaya, Mahirogullari: "Chondrocyte proliferation, viability and differentiation is declined following administration of methylphenidate utilized for the treatment of attention-deficit/hyperactivity disorder." in: Human & experimental toxicology, (2016) (PubMed).

Wu, Ng, Chen, Steer, Song: "MicroRNA-21 is a potential link between non-alcoholic fatty liver disease and hepatocellular carcinoma via modulation of the HBP1-p53-Srebp1c pathway." in: Gut, (2015) (PubMed).

Dogan, Isyar, Yilmaz, Bilir, Sirin, Cakmak, Mahirogullari: "Are the leading drugs against Staphylococcus aureus really toxic to cartilage?" in: Journal of infection and public health, (2015) (PubMed).

Zhang, Ge, Fuchs: "miR-125b can enhance skin tumor initiation and promote malignant progression by repressing differentiation and prolonging cell survival." in: Genes & development, Vol. 28, Issue 22, pp. 2532-46, (2014) (PubMed).

Ren, Wang, Jiang, Dai, Jiang: "Anti-tumor effect of a novel soluble recombinant human endostatin: administered as a single agent or in combination with chemotherapy agents in mouse tumor models." in: PLoS ONE, Vol. 9, Issue 9, pp. e107823, (2014) (PubMed).

Antigendetails

Hintergrund: The measurement and monitoring of cell proliferation is an essential technique in any laboratory focused on cell-based research. This skill allows for the optimization of cell culture conditions as well as the determination of cytokine, growth factor, or hormone activity. More importantly, the cytostatic nature of anticancer compounds in toxicology testing, the efficacy of therapeutic chemicals in drug screening, and cell-mediated cytotoxicity can all be assessed through the quantification and monitoring of cell proliferation. Cell proliferation characteristics include cellular metabolic activity and cell membrane integrity. One method for measuring metabolic activity is to incubate the cells with a tetrazolium salt such as WST-1, which is cleaved into a colored formazan product by metabolically active cells. Similarly, the green fluorescent dye Calcein AM can measure intracellular esterase activity in proliferating live cells, which is another indicator of cell viability.