A6885

Manufactured by Merck Group

Sourced in United Kingdom, Sao Tome and Principe

A6885 is a laboratory equipment product from Merck Group. It serves as a core function to support scientific research and testing activities.

Automatically generated - may contain errors

Lab products found in correlation

A8960, by Merck Group (2 mentions) Neural induction supplement, by Thermo Fisher Scientific (1 mentions) Brain derived neurotrophic factor (bdnf), by Thermo Fisher Scientific (1 mentions) Y0503, by Merck Group (1 mentions) Glutamax, by Thermo Fisher Scientific (1 mentions) Y 27632, by Merck Group (1 mentions) L ascorbic acid, by Merck Group (1 mentions) A8380, by Merck Group (1 mentions) R2625 50mg, by Merck Group (1 mentions) Pt 1200e, by Kinematica (1 mentions) Phosstop, by Roche (1 mentions) Poly l ornithine, by Merck Group (1 mentions)

5 protocols using a6885

Differentiation of iPSCs into Functional Neurons

Cited 1 time

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NPC1 iPSCs were differentiated into neural stem cells (NSCs) by using a kit from Life Technologies. Briefly, iPSCs were digested with Dispase and re-seeded onto Geltrex-coated plate at 20% confluence. After cells attached, the medium was changed to Induction Medium containing the Neurobasal Medium plus Neural Induction Supplement (A15640SA, Life Technologies). At day 7 of neural induction, the initial NSCs were dissociated with Accutase and plated for further expansion in Neural Expansion Medium containing Neurobasal Medium and Advanced DMEM/F12 with 1x Neural Induction Supplement. NSCs were characterized by staining with antibodies against nestin and Sox2 that showed 99% positive cells with both neural markers.
For neuronal differentiation, dissociated NSCs were cultured on poly-L-ornithine and laminin coated 96-well plates in the Induction Medium with 5 μM Y27632, a ROCK inhibitor (Y0503, Sigma-Aldrich) for one day. The medium was changed to a differentiation medium containing Neurobasal Medium, 1x B27 (17504-044, Life Technologies), 1x glutamax (35050, Life Technologies), 200μM L-ascorbic acid (A8960, Sigma-Aldrich), 1μM cAMP (A6885, Sigma-Aldrich), 10ng/ml BDNF (10908-010, Life Technologies) and 10ng/ml GDNF (PHC7044, Life Technologies) that was changed every two days for 12 days.^{20 (link)}

Yu D., Swaroop M., Wang M., Baxa U., Yang R., Yan Y., Coksaygan T., DeTolla L., Marugan J.J., Austin C.P., McKew J.C., Gong D.W, & Zheng W. (2014). Niemann-Pick disease type C: induced pluripotent stem cell-derived neuronal cells for modeling neural disease and evaluating drug efficacy. Journal of biomolecular screening, 19(8), 1164-1173.

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Endometrial Stromal Cell Culture and Treatment

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Primary endometrial stromal cells (ESCs) were isolated from endometrial biopsies as previously described [4 (link)], yielding a ~ 95% pure stromal cell culture [13 (link)]. ESCs (6 × 10⁵ cells) were cultured in 6-well plates for 24 h prior to treatments. Confluent ESC monolayers were treated with cAMP (500 μM, A6885, Sigma, UK), DHT (10⁻⁸ M, A8380, Sigma), and DHT + cAMP (10⁻⁸ M + 500 μM) for 24 h, 48 h, and 72 h. RNA was extracted after treatments and cell morphology assessed throughout the duration of the experiment.

Younas K., Quintela M., Thomas S., Garcia-Parra J., Blake L., Whiteland H., Bunkheila A., Francis L.W., Margarit L., Gonzalez D, & Conlan R.S. (2019). Delayed endometrial decidualisation in polycystic ovary syndrome; the role of AR-MAGEA11. Journal of Molecular Medicine (Berlin, Germany), 97(9), 1315-1327.

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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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Muscle Homogenate Preparation and cAMP Treatment

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Whole tissue homogenate samples were prepared by homogenizing ~5 mg of frozen EDL muscle (1:50 wt/vol) using a handheld polytron homogenizer (Polytron PT 1200E Kinematica, Lucerne, Switzerland) three times at maximum speed for 5 to 10 s in the ice-cold homogenizing solution (50 mM HDTA, 90 mM HEPES, 126 mM K⁺, 36 mM Na⁺, 8 mM ATP, 8.5 mM total Mg²⁺, 10 mM creatine phosphate (pH 7.10) with phosphatase inhibitor (PhosSTOP; Roche Diagnostics, Mannheim, Germany), 295 ± 10 mosmol/kg H₂O). Following homogenization, whole muscle samples were vortexed three times at maximum speed for ~10 s and then diluted to 10 µg/µL in homogenizing solution. The homogenate samples were then divided into three tubes (50 to 100 µL each) for control and cAMP-treatments (cAMP). We performed treatments using sodium-salt cAMP (Sigma A6885, same as fiber Ca²⁺ experiments). For cAMP treatment, cAMP was added to the whole muscle homogenates at 100 µM, using a stock solution (dissolved in ddH₂O). Samples were vortexed for ~10 s then left for 1 min at room temperature (~23 °C). After 1 min, 3XSDS loading buffer was added (2:1 v/v) to all samples, which were then incubated at RT for ~1 h and stored at −80 °C until Western blot analysis.

Singh D.P., Pearce L., Choi R.H., Meizoso-Huesca A., Wette S.G., Scott J.W., Lamboley C.R., Murphy R.M, & Launikonis B.S. (2023). Evolutionary isolation of ryanodine receptor isoform 1 for muscle-based thermogenesis in mammals. Proceedings of the National Academy of Sciences of the United States of America, 120(4), e2117503120.

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Differentiation of Neural Progenitor Cells

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The neurons were differentiated according to Zhang et al. (2017 (link)). When the NPCs reached 70% confluency, they were plated on Matrigel in a neural expansion media (NEM). The NEM was supplemented with 20 ng/ml of bFGF, and EGF. The NPCs were re-plated several times until passage three. Then, they (for last passage) were plated on 0.001% Poly-L-Ornithine (P4957; Sigma-Aldrich, St. Louis, MO, United States) and a 5 μg/ml laminin (L2020; Sigma-Aldrich, St. Louis, MO, United States) coating in a neural maturation media (NMM) for 10 days. The NMM was supplemented with 20 ng/μl brain-derived neurotrophic factor (BDNF) (CYT-207; Prospec, Rehovot, Israel), 10 ng/μl glial cell line-derived neurotrophic factor (GDNF) (CYT-305; Prospec, Rehovot, Israel), 200 μM L-ascorbic acid 2-phosphate (A8960, Sigma-Aldrich, St. Louis, MO, United States), and 50 μM dibutyryl-cyclic adenosine monophosphate (db-cAMP) (A6885; Sigma-Aldrich, St. Louis, MO, United States). Half media change was performed every 3 days. Ten-day immature neurons were passaged a last time on PLO-Laminin coated plates with the NMM at a seeding density of 50,000 cells/cm² until they reached maturation for the metabolic assays.

Salcedo C., Andersen J.V., Vinten K.T., Pinborg L.H., Waagepetersen H.S., Freude K.K, & Aldana B.I. (2021). Functional Metabolic Mapping Reveals Highly Active Branched-Chain Amino Acid Metabolism in Human Astrocytes, Which Is Impaired in iPSC-Derived Astrocytes in Alzheimer's Disease. Frontiers in Aging Neuroscience, 13, 736580.

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