R2625 50mg

Manufactured by Merck Group

R2625-50MG is a laboratory reagent manufactured by Merck Group. It is a powdered form of a chemical compound used in various research and analytical applications. The core function of this product is to serve as a research tool for scientific experimentation and analysis, without any specific interpretation or extrapolation on its intended use.

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Lab products found in correlation

A6885, by Merck Group (1 mentions) Cabd353003, by Merck Group (1 mentions) Trypsin edta, by Thermo Fisher Scientific (1 mentions)

3 protocols using r2625 50mg

Retinoic Acid and cAMP-Induced HT22 Cell Differentiation

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Cultured HT22 cells
were differentiated by applying 10 μM retinoic acid (Sigma-Aldrich,
R2625-50MG) and 500 μM cyclic adenosine monophosphate (cAMP,
Sigma-Aldrich, A6885) in DMEM medium supplied with 0.5% FBS (differentiation
medium) and incubated for 48 h before adding other treatments. L-Glutamate
acid at 5 mM was applied for 24 h of incubation. Then, the 2′-7′dichlorofluorescin
diacetate (DCFH-DA) fluorescent probe was added to cells in DMEM-only
medium at 10 μM working concentration for a total of 30 min
incubation with gentle shaking every 5 min at 37 °C. After removing
the fluorescent probe, the cells were washed with probe-free DMEM-only
medium three times to wash away redundant probe. Tested compounds
(cA, cB, cC: 2 μg/mL,
in molar units, cA: 3.23, cB: 3.15, cC: 3.13 μM), other control drugs (memantine, nimodipine,
DB1246: 2 μM), and positive ROS control (Rosup: 50 μg/mL)
were then applied to different wells of cells for 1 h of incubation.
The fluorescence intensity of dichlorofluorescein (DCF), oxidated
from DCFH by cellular ROS, was then measured using a microplate reader
with 488/525 nm excitation/emission filters.

Cheng A., Liu C., Ye W., Huang D., She W., Liu X., Fung C.P., Xu N., Suen M.C., Ye W., Sung H.H., Williams I.D., Zhu G, & Qian P.Y. (2022). Selective C9orf72 G-Quadruplex-Binding Small Molecules Ameliorate Pathological Signatures of ALS/FTD Models. Journal of Medicinal Chemistry, 65(19), 12825-12837.

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Retinoic Acid-Induced Differentiation

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Differentiation was induced by seeding cells at a density of 0.3×10⁶ cells per well in 6-well plates (for ATAC-seq) or at 1.5×10⁶ cells per 10-cm culture dish (for ChIP-seq) in media containing 1 μM all-trans retinoic acid (RA,Sigma-Aldrich R2625–50MG) and no LIF. Cells were differentiated for 48 hours and the media was changed after 24 hours. Control cells were treated with a DMSO volume equivalent to the RA volume in the differentiating cells. After 48 hours, the cells were harvested, washed once with PBS, counted and used immediately in either ChIP-seq or ATAC-seq experiments.

Uckelmann M., Levina V., Taveneau C., Han Ng X., Pandey V., Martinez J., Mendiratta S., Houx J., Boudes M., Venugopal H., Trépout S., Zhang Q., Flanigan S., Li M., Sierecki E., Gambin Y., Das P.P., Bell O., de Marco A, & Davidovich C. (2024). Dynamic PRC1-CBX8 stabilizes a porous structure of chromatin condensates. bioRxiv.

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Quantifying Pluripotency Dynamics in mESCs

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Differentiation analysis was performed as in Pucci et al. (2013) (link). Briefly, after 2 days of growth in LIF-containing media (as above), cells were plated at a density of approximately 0.2 × 10⁶ cells/100 mm plate on non-gelatin-coated dishes (VWR, cat. # CABD353003), in media without LIF but supplemented with 5 μM all-trans retinoic acid (ATRA, Sigma, R2625-50MG) for 6 days. At day 6, cells were trypsinized using 0.05% w/v trypsin-EDTA 0.5 mM (Invitrogen, cat. # 25300–054), transferred to gelatin-coated plates in GMEM media without ATRA and supplemented with LIF (as above) for 6 days or longer, as indicated. Cells were split to ensure they did not exceed 80% confluency After each step of the differentiation assay, and before reseeding the trypsinized cells, 0.2–0.5 × 10⁶ cells were used for flow cytometry analysis or 3 × 10⁶ cells for FACS. The pluripotent state was assessed by flow cytometry, using the Pou5f1-GFP reporter gene as a quantitative tool (see below). All experiments were repeated at least three times (n = 3 biological replicates), and typically included n = 3 technical replicates. A biological replicate represents a distinct mESC population (usually analyzed on a different day), and a technical replicate is the same biological replicate processed separately (usually on the same day).

Criqui M., Qamra A., Chu T.W., Sharma M., Tsao J., Henry D.A., Barsyte-Lovejoy D., Arrowsmith C.H., Winegarden N., Lupien M, & Harrington L. (2020). Telomere dysfunction cooperates with epigenetic alterations to impair murine embryonic stem cell fate commitment. eLife, 9, e47333.

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