Neutral red

Manufactured by Agilent Technologies

Neutral red is a vital dye used in laboratory settings. It is a reddish-orange crystalline compound that exhibits a pH-dependent color change, making it useful for various applications in biological and chemical analysis.

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Lab products found in correlation

Glacial acetic acid, by Merck Group (2 mentions) Neutral red, by Merck Group (2 mentions) Microplate reader, by Agilent Technologies (1 mentions) Microplate spectrophotometer, by Agilent Technologies (1 mentions)

2 protocols using neutral red

Neutral Red-Based Cell Growth Assay

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After transfection, cells were incubated for 5 d. The Idh2 knockdown cells were re-plated on a 12-well plate. Cells were allowed to grow from 24 to 96 h in DMEM. Cell growth rates were determined by the neutral red assay as previously described (Repetto et al., 2008). At completion of incubation, cells were incubated in DMEM with 50 µg/mL of neutral red (Sigma-Aldrich, St. Louis, MO) at 37 °C for 2–3 h. After washing, cells were then treated with 200 μl of neutral red solubilization solution (50% ethanol, 49% deionized water, and 1% glacial acetic acid (Sigma-Aldrich, St. Louis, MO) per well. The 12-well plate was incubated at room temperature on a plate shaker overnight. The OD (optical density) values of the neutral red extract in each well were measured at 540 nm in a microplate reader (Bio-Tek).

White K., Kim M.J., Han C., Park H.J., Ding D., Boyd K., Walker L., Linser P., Meneses Z., Slade C., Hirst J., Santostefano K., Terada N., Miyakawa T., Tanokura M., Salvi R, & Someya S. (2018). Loss of IDH2 Accelerates Age-related Hearing Loss in Male Mice. Scientific Reports, 8, 5039.

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Hydrogen Peroxide Stress Assay for Sirt1 Knockdown

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The Sirt1 knockdown or control cells were replated on a 96 well plate (3X10⁴/well) and treated with hydrogen peroxide at 0 to 2.8 mM for 2 hours. For cell viability measurements, after 22 hours, the media was replaced with DMEM containing 50 μg/mL neutral red (Sigma-Aldrich, St. Louis, MO) as previously described (Someya, et al., 2009 (link)). After 2 hours, 200 μl of a neutral red destaining solution composed of 50% ethanol, 49% deionized water, and 1% glacial acetic acid (Sigma-Aldrich, St. Louis, MO) was added to each well. The 96-well plate was placed on a plate shaker for 1 hour and the OD of the neutral red extract in each well was measured at 540 nm in a microplate spectrophotometer (BioTek, Winooski, VT). Each condition was run in duplicate.

Han C., Linser P., Park H.J., Kim M.J., White K., Vann J.M., Ding D., Prolla T.A, & Someya S. (2016). Sirt1 deficiency protects cochlear cells and delays the early onset of age-related hearing loss in C57BL/6 mice. Neurobiology of aging, 43, 58-71.

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