Six epigenetic modifiers were opted based on literature review viz., two DNMT inhibitors 5-azacytidine (A2385, Sigma-Aldrich) and hydralazine hydrochloride (H1753, Sigma-Aldrich), a sirtuins activator [quercetin (Q4951, Sigma-Aldrich)] and three HDAC inhibitors Suberoylanilide Hydroxamic Acid (SAHA) (390585, Sigma-Aldrich), sodium butyrate (303410, Sigma-Aldrich) and valproic acid (P4543, Sigma-Aldrich)] (de la Cruz et al., 2012 (link); González-Menéndez et al., 2016 (link)).
A2385
Manufactured by Merck Group
Sourced in United States
A2385 is a laboratory instrument designed for the measurement and analysis of various samples. It is capable of performing accurate and precise measurements, but a detailed description of its core function cannot be provided while maintaining an unbiased and factual approach. The intended use of this product may vary depending on the specific requirements of the user.
Automatically generated - may contain errors
Lab products found in correlation
Valproic acid, by Merck Group (1 mentions)
Suberoylanilide hydroxamic acid saha, by Merck Group (1 mentions)
P4543, by Merck Group (1 mentions)
Q4951, by Merck Group (1 mentions)
5 azacytidine, by Merck Group (1 mentions)
Quercetin, by Merck Group (1 mentions)
Sodium butyrate, by Merck Group (1 mentions)
Ab52862, by Abcam (1 mentions)
A7811, by Merck Group (1 mentions)
A5960, by Merck Group (1 mentions)
6 protocols using a2385
1
Epigenetic Modifiers Protocol
2
Prenatal Exposure: Epigenetic Impacts
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Ninety day old female Fisher inbred strain rat dams were kept overnight with males. If the sperm was found in the vaginal smear the next morning, it designated gestation day (GD) 0. On GD 12–13, pregnant dams received 5azaC (5 mg/kg, A 2385, Sigma, St. Louis, MO, USA) dissolved in phosphate buffered saline (PBS) by an i.p. injection. PBN (40 mg/kg, B 7263, Sigma, St. Louis, MO, USA) was injected in the tail vein as pretreatment 1 h before 5azaC or alone. A total of 352 placentas were isolated from four groups of dams i.e., control, PBN, 5azaC, and PBN + 5azaC-treated pregnant dams, 177 at GD 15 and 175 at GD 20.
3
Anther Culture with Cold and Abscisic Acid
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Tillers were harvested when most of microspores were at mid- to late-uninucleate development stage, then wrapped in plastic bags, placed in jars containing Hoagland’s salt solution (HSS, according to Wędzony58 ) and stored at 4 °C in the dark for 3 weeks. AC (Sigma-Aldrich; A2385) was added to HSS at a concentration of 2.5, 5.0 and 10 μM for the last 4 days before anther isolation. The solution was refreshed three times every 24 h. After control (low temperature) and combined (low temperature with AC) tillers treatments, the anthers were isolated and transferred to in vitro culture or collected for proteomic and molecular analyses (immediately frozen in liquid nitrogen and stored at -80 °C).
4
Preparation of B[a]P Dispersion Medium
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B[a]P was purchased from (A2385, Sigma Aldrich Co., LLC, St. Louis, MO, USA), dissolved in a modified dispersion medium (DM) of 5.5 mM D-glucose and 0.01 mg/l 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) in Ca2+ and Mg2+-free phosphate buffer saline (PBS), sonicated by a bath-type sonicator, and stored in a refrigerator at 4 °C. Bovine serum albumin (BSA) was removed from the original DM, which was developed for the dispersion of nanoparticles and described previously [34 (link),35 (link)], to prepare the modified DM to optimize the dispersion of B[a]P.
5
Evaluating NAFLD Treatments in Mice
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Male C57BL6/J mice aged 4 to 5 weeks and weighing 18 to 20 g were kept on a combination diet of high fat and high fructose for 10 weeks, whereas control mice were kept on normal pellet diet (3.8 kcal/g) (Normal Control). Alteration in the metabolic parameters such as an increase in plasma insulin, glucose, triglycerides, and circulating lipids indicates successful NAFLD induction. It was followed by the randomization of animals into four groups, which were administered treatment for 8 weeks:
Group 1: Normal Control; received 0.9% w/v normal saline (CON); n = 6,
Group 2: NAFLD Disease Control; received a combination diet of high fat and high fructose (HF-HFR); n = 6,
Group 3: NAFLD mice administered Aza (0.25 mg/kg i.p., NAFLD + Aza); n = 6.
Group 4: NAFLD mice administered Zeb (200 mg mg/kg i.p., NAFLD + Zeb); n = 6.
The dose of Zeb (TCI # Z0022) and Aza (Sigma # A2385) used for animal studies was taken from previous studies, and at these doses both agents were considered safe (22 , 23 (link)).
Group 1: Normal Control; received 0.9% w/v normal saline (CON); n = 6,
Group 2: NAFLD Disease Control; received a combination diet of high fat and high fructose (HF-HFR); n = 6,
Group 3: NAFLD mice administered Aza (0.25 mg/kg i.p., NAFLD + Aza); n = 6.
Group 4: NAFLD mice administered Zeb (200 mg mg/kg i.p., NAFLD + Zeb); n = 6.
The dose of Zeb (TCI # Z0022) and Aza (Sigma # A2385) used for animal studies was taken from previous studies, and at these doses both agents were considered safe (22 , 23 (link)).
6
hCMPCs Cardiomyocyte Differentiation Protocol
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hCMPCs were induced to differentiate into cardiomyocytes according to the protocol described previously [17] (link). First, 5 mM 5-azacytidine (A2385, Sigma, USA) was added for three days; then, hCMPCs were treated with transforming growth factor (TGF) -β1 (1 ng/ml, 100-21c, PeproTech, USA) and ascorbic acid (100 µM, A5960, Sigma, USA). After treatment for 14 days, RNA was extracted for RT-PCR. We detected the expression of transcription factors indicative of cardiomyocyte predisposition (MEF2C, GATA-4, Nkx-2.5) and genes encoding sarcomere assembly proteins (α-Actinin, β-MHC). In addition, sarcomeric proteins α-Actin (A7811, Sigma, USA) and Troponin I (ab52862, Abcam, USA) were stained to quantify the degree of differentiation based on a previously provided protocol [17] (link). The cells were photographed with the Live Cell Imaging System (Leica AF 7000, Leica).
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