Alda 1

Manufactured by Merck Group

Sourced in United States, Germany

Alda-1 is a laboratory equipment product manufactured by Merck Group. It is designed for general laboratory use. The core function of Alda-1 is to provide a reliable and consistent platform for various laboratory procedures and experiments.

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Lab products found in correlation

Daidzin, by Merck Group (3 mentions) Dimethyl sulfoxide (dmso), by Merck Group (2 mentions) Monocrotaline mct, by Merck Group (1 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions) Amicon ultra, by Merck Group (1 mentions) Spectramax 340pc, by Molecular Devices (1 mentions) Necrosulfonamide nsa, by Abcam (1 mentions) Lipopolysaccharide lps, by Merck Group (1 mentions) Atractyloside, by Merck Group (1 mentions) Superoxide dismutase sod assay kit, by Nanjing Jiancheng (1 mentions) Anti bcl 2, by Santa Cruz Biotechnology (1 mentions) Mouse anti aldh2, by Santa Cruz Biotechnology (1 mentions) Anti β actin monoclonal antibody, by Santa Cruz Biotechnology (1 mentions) Anti bax, by Santa Cruz Biotechnology (1 mentions) Streptozotocin (stz), by Merck Group (1 mentions) Lps from escherichia coli o55 b5, by Merck Group (1 mentions) Gapdh antibody, by Boster Bio (1 mentions) Nec 1, by Merck Group (1 mentions) Cleaved caspase 3 antibody, by Cell Signaling Technology (1 mentions) Mitochondrial aldh2 activity assay kit, by Abcam (1 mentions)

19 protocols using alda 1

Alda-1 Attenuates High Glucose-Induced Cardiac Fibroblast Injury

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CFs were divided into 7 groups after incubated (37°C, 5% CO₂) in a serum-free DMEM medium for 48 h:

Group 1: Normal group (NG), CFs were cultured with DMEM medium with normal glucose (glucose concentration at 5.5 mM) and treated with the same volume of solvent instead of drug.

Group 2: Normal glucose group + Alda-1 (NG + Alda-1), Alda-1 at 20 μM (the specific agonist of ALDH2, Sigma-Aldrich Co., St. Louis, MO, USA) [14 (link)] was added into DMEM medium with normal glucose and cultured for 48 h.

Group 3: High glucose groups (HG), CFs were cultured in DMEM medium with high glucose (glucose concentration at 30 mM) to induce injury for 48 h.

Group 4: HG + Alda-1 group (HG + Alda-1), for observing whether activation of ALDH2 can attenuate HG-induced CF injury, 20 μM Alda-1 was added into DMEM medium with 30 mM glucose and cultured for 48 h.

Group 5: HG + Alda-1 + daidzin (HG + Alda-1 + daidzin), 20 μM Alda-1 and 60 μM daidzin (the specific antagonist of ALDH2, Sigma-Aldrich Co., St. Louis, MO, USA) [15 (link)] were added into DMEM medium with 30 mM glucose and cultured for 48 h.

Group 6: HG + daidzin group (HG + daidzin), 60 μM daidzin was added into DMEM medium with 30 mM glucose and cultured for 48 h.

Group 7: Hypertonic group (HPG), for excluding the role of hypertonic, CFs were cultured with DMEM medium with 5.5 mM glucose and treated with 24.5 mM mannitol 48 h.

Gu X., Fang T., Kang P., Hu J., Yu Y., Li Z., Cheng X, & Gao Q. (2017). Effect of ALDH2 on High Glucose-Induced Cardiac Fibroblast Oxidative Stress, Apoptosis, and Fibrosis. Oxidative Medicine and Cellular Longevity, 2017, 9257967.

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Alda-1 Ameliorates Monocrotaline-Induced PAH

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All animal care and experimental procedures were approved and conducted in accordance with the Institutional Animal Care and Use Committee of Jinzhou Medical University and conformed to the Guide for the Care and Use of Laboratory Animal published by the US National Institutes of Health. Male Sprague–Dawley rats (n=48; weighing 220–250 g) were purchased from Vital River Laboratories Animal Company (Beijing, China). The animals were intraperitoneally (i.p.) injected with a single dose of monocrotaline (MCT; 60 mg/kg; Sigma-Aldrich, St. Louis, MO) to induce severe PAH within 2 or 4 weeks (n=8 each group). For experiments involving pre-treatment with Alda-1 (Sigma-Aldrich Co., St. Louis, MO), MCT-injected rats were randomly divided into 3 groups, including the MCT group (n=12), the vehicle-alone group (n=6) administered 50% polyethylene glycol (PEG) and 50% dimethyl sulfoxide (DMSO) by volume, and the Alda-1 group (n=6). Control rats (n=8) were injected with an equal volume of 0.9% phosphate-buffered saline (PBS). The MCT-treated rats were subcutaneously implanted with mini-osmotic pumps (model 2004; ALZET, Cupertino, CA) and continuously infused with Alda-1 (10 mg kg⁻¹ d⁻¹) for 4 weeks.

Xu T., Liu S., Ma T., Jia Z., Zhang Z, & Wang A. (2016). Aldehyde dehydrogenase 2 protects against oxidative stress associated with pulmonary arterial hypertension. Redox Biology, 11, 286-296.

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Modulating ALDH2 for Cardioprotection

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H9C2 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) (Gibco, Grand Island, NY, USA) supplemented with 10% (v/v) fetal bovine serum (FBS) and 1% penicillin/streptomycin. All cells were maintained in a humidified incubator with 95% air/5% CO² at 37°C. For in vitro study, H9C2 cells were incubated with the ALDH2 activator Alda-1 (20 μM, Sigma, USA) or ALDH2 inhibitor Daidzin (60 μM, Sigma, USA) for 30 min at 37°C (95% air/5% CO²) before a 120 min exposure to hypoxia (1% air/5% CO²/94% N²), followed by 60 min reoxygenation (95% air/5% CO²). Furthermore, H9C2 cells were treated with H/R in the absence or presence of Alda-1 with the mitophagy inducer carbonyl cyanide chlorophenylhydrazone (CCCP) (10 μM, Sigma, USA) to evaluate whether and how ALDH2 could affect mitophagy in cardiomyocytes.

Ji W., Wei S., Hao P., Xing J., Yuan Q., Wang J., Xu F, & Chen Y. (2016). Aldehyde Dehydrogenase 2 Has Cardioprotective Effects on Myocardial Ischaemia/Reperfusion Injury via Suppressing Mitophagy. Frontiers in Pharmacology, 7, 101.

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Alda-1 compound protocol

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Alda-1 (N-(1,3-Benzodioxol-5-ylmethyl)-2,6-dichlorobenzamide) (St. Louis, MO) was purchased from Sigma Aldrich, protected from light, and 20µM concentrations were used for experiments.

Patil S.S., Hernández-Cuervo H., Fukumoto J., Narala V.R., Saji S., Borra M., Alleyn M., Lin M., Soundararajan R., Lockey R., Kolliputi N, & Galam L. (2019). Alda-1 attenuates hyperoxia-induced mitochondrial dysfunction in lung vascular endothelial cells. Aging (Albany NY), 11(12), 3909-3918.

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Quantifying AcH Metabolism Kinetics in ESCs

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We cultured ESCs in either 6-well plates or 10 cm dishes and harvested in TEN buffer pH=7.2 (50 mM Tris, 150 mM NaCl, and 1 mM EDTA) to retain enzymatic activity of cells. We then centrifuged the lysates, resuspended them in 250 mM Tris pH=7.8 with vortexing, and lysed them by performing snap thawing after freezing at −70°C. The ESC lysates were further concentrated using 10K concentrators (Amicon Ultra, Sigma), and by performing two additional concentration steps using 250 mM Tris pH=7.8. Kinetics assays were conducted to determine the rate of AcH metabolism in various cell lines using the SpectraMAX 340PC plate reader (Molecular Devices, San Jose, CA). 0.99 mg/ml of protein lysate was incubated in PBS buffer (pH 7.4) supplemented with NAD⁺ 3 mM, Alda1 (Sigma) 15 μM, or diethylbenzaldehyde (DEAB) (Sigma) 1 μM. After three minutes, the assay was initiated by the addition of 2.7 mM AcH. We monitored the absorbance every 60 seconds at 340 nm for NADH formation and at 600 nm for background determination. The averages of three technical replicates for each sample were used to quantitate rate values in nmol•min⁻¹•g⁻¹ protein units, and two biological repeats were performed. Rates were calculated using the slopes of linear regression lines generated for the average of each sample in Microsoft Excel.

Serio R.N., Lu C., Gross S.S, & Gudas L.J. (2019). Different effects of knockouts in ALDH2 and ACSS2 on embryonic stem cell differentiation. Alcoholism, clinical and experimental research, 43(9), 1859-1871.

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Aldh2 Knockout Mice Model of LPS-Induced Injury

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Six to 8 weeks old C57BL/6J mice, Aldh2 knockout mice (Aldh2^−/−) were used. Only male mice were chosen for experiment to avoid sex hormone interference. Mice were housed in an appropriate environment (23.0°C ± 2.0°C, 45%–50% humidity) with a 12/12-light/dark cycle, and they had access to food and water ad libitum until experimentation. They were randomly divided into different groups. Model was established by administrating 15 mg/kg of Lipopolysaccharide (LPS, Sigma) intraperitoneally (i.p.) 12 h before sacrifice. Alda-1 (20 mg/kg i.p., Sigma-Aldrich) or necrosulfonamide (NSA, 20 mg/kg i.p., Abcam) was administered half an hour before LPS injection. An equal amount of pathogen-free normal saline (NS) was used as control.

Zhang Y., Lv Y., Zhang Q., Wang X., Han Q., Liang Y., He S., Yuan Q., Zheng J., Xu C., Zhang X., Wang Z., Yu H., Xue L., Wang J., Xu F., Pang J, & Chen Y. (2023). ALDH2 attenuates myocardial pyroptosis through breaking down Mitochondrion-NLRP3 inflammasome pathway in septic shock. Frontiers in Pharmacology, 14, 1125866.

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Antioxidant and Apoptosis Regulation in Cell Model

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Streptozotocin (STZ), cyanamide (CYA), wortmannin (Wor), atractyloside (Atr), and Alda-1 were purchased from Sigma (St. Louis, MO, USA). Ethanol (EtOH) was obtained from Bengbu New Chemical Reagent Factory, China. 10% fetal bovine serum was obtained from Zhe Jiang Tianhang Biological corporation, China. Lactate dehydrogenase (LDH), Malondialdehyde (MDA), and superoxide dismutase (SOD) assay kits were purchased from Nanjing Jiancheng Bioengineering Institute, China. CCK-8 assay kit was from Shanghai Bestbio Life Technology, China. The primers used were as follows: for ALDH2 forward: 5′-GTG TTC GGA GAC GTC AAA GA-3′ and reverse 5′-GCA GAG CTT GGG ACA GGT AA-3′, the amplified fragment length was 187 bp; for Bcl-2 forward: 5′-CTG GTG GAC AAC ATC GCT CTG-3′ and reverse: 5′-GGT CTG CTG ACC TCA CTT GTG-3′, the amplified fragment length was 227 bp; for Bax forward: 5′-GGA TCG AGC AGA GAG GAT GG-3′ and reverse: 5′-GCT CAT TGC CGA TAG TGA TGA CT-3′, the amplified fragment length was 464 bp; for β-actin forward: 5′-GAT GGT GGG TAT GGG TCA GAA-3′ and reverse: 5′-GGC CAT CTC TTG CTC GAA GTC-3′, the amplified fragment length was 630 bp. Mouse anti-ALDH2, anti-Bcl-2, anti-Bax, and anti-β-actin monoclonal antibodies were purchased from Santa Cruz Biotechnology (CA). Goat anti-mouse secondary antibodies were from Boston Co., Ltd., Wuhan, China.

Kang P.F., Wu W.J., Tang Y., Xuan L., Guan S.D., Tang B., Zhang H., Gao Q, & Wang H.J. (2016). Activation of ALDH2 with Low Concentration of Ethanol Attenuates Myocardial Ischemia/Reperfusion Injury in Diabetes Rat Model. Oxidative Medicine and Cellular Longevity, 2016, 6190504.

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Cardioprotective effects of ALDH2 modulation

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Rat cardiomyoblasts cell line (H9c2) and primary cardiomyocytes isolated from adult male wild type (WT) mice and ALDH2 knockout (KO) mice were used. Cells were cultured normally in a humidified incubator with 95% air/5% CO₂ at 37°C. Hypoxia/reoxygenation procedure was induced by exposing cells in a hypoxic workstation (H35, Don Whitley Scientific, Bingley, United Kingdom) containing 94% N₂, 5% CO₂, and 1% O₂ at 37°C with serum-deprived media for 4 h and then culturing cells under normal conditions with complete media for another 2 h. Alda-1 (20 mmol/L) or Daidzin (60 mmol/L, Sigma-Aldrich) was added to the media 30 min before hypoxia/reoxygenation.

Zhang R., Liu B., Fan X., Wang W., Xu T., Wei S., Zheng W., Yuan Q., Gao L., Yin X., Zheng B., Zhang C., Zhang S., Yang K., Xue M., Wang S., Xu F., Wang J., Cao Y, & Chen Y. (2020). Aldehyde Dehydrogenase 2 Protects Against Post-Cardiac Arrest Myocardial Dysfunction Through a Novel Mechanism of Suppressing Mitochondrial Reactive Oxygen Species Production. Frontiers in Pharmacology, 11, 373.

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Activating ALDH2 in Cardiomyocytes

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To activate ALDH2, cardiomyocytes were treated with Alda-1 (20 μM, Sigma-Aldrich, St. Louis, MO, USA) for 12 h [19 (link),30 (link),31 (link)]. ALDH2 activity was detected by the assay kit from GENMED SCIENTIFICS. Normal and ALDH2-activated mitochondria were collected for further analyses.

Sun X., Gao R., Li W., Zhao Y., Yang H., Chen H., Jiang H., Dong Z., Hu J., Liu J., Zou Y., Sun A, & Ge J. (2021). Alda-1 treatment promotes the therapeutic effect of mitochondrial transplantation for myocardial ischemia-reperfusion injury. Bioactive Materials, 6(7), 2058-2069.

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ALDH2 Activation Modulates Sepsis Pathogenesis

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Eight to ten weeks old C57BL/6 (wild-type (WT)) mice, ALDH2^−/− mice, and their littermates weighing 20–25 g were used in our experiments. The C57BL/6 mice were obtained from the Shanghai Animal Administration Center (Shanghai, China), and the ALDH2^−/− mice were provided by Professor Sun (Fudan University). All the mice were housed in a climate-controlled environment with free access to food and water ad libitum. The sepsis model was induced via intraperitoneal (IP) injection of lipopolysaccharide (LPS) from Escherichia coli O55:B5 (Sigma, St. Louis, MO, USA) at a dose of 10 mg/kg. Alda-1 (Sigma, St. Louis, MO, USA), an ALDH2-specific activator, was administered via IP injection at a dose of 10 mg/kg 1 hr before LPS administration. An IP injection of an equal volume of saline or DMSO (Sigma) served as vehicle-treated groups. The mice were randomized into the following groups: WT, ALDH2 knockout (KO), WT + LPS (10 mg/kg, IP), KO + LPS (10 mg/kg, IP), WT + Alda-1 (10 mg/kg, IP), and WT + Alda-1 + LPS (10 mg/kg, respectively, IP). All animal study protocols were approved by the Animal Care and Use Committee of Fudan University (Shanghai, China). All animal procedures followed the National Institutes of Health's guidelines for the care and use of laboratory animals.

Jin J., Chang R.S., Xu S., Xia G., Wong J.M., Fang Y., Jia P, & Ding X. (2023). Aldehyde Dehydrogenase 2 Ameliorates LPS-Induced Acute Kidney Injury through Detoxification of 4-HNE and Suppression of the MAPK Pathway. Journal of Immunology Research, 2023, 5513507.

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