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4 2 aminoethyl benzenesulfonyl fluoride hydrochloride aebsf

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4-(2-Aminoethyl) benzenesulfonyl fluoride hydrochloride (AEBSF) is a serine protease inhibitor. It is commonly used in laboratory settings to inactivate proteases during protein purification or cell lysis procedures.

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Lab products found in correlation

Pepstatin a, by Merck Group (2 mentions) Cycloheximide (chx), by Merck Group (1 mentions) Tunicamycin, by Merck Group (1 mentions) Cip2a, by Cell Signaling Technology (1 mentions) Gapdh, by Cell Signaling Technology (1 mentions) β actin, by Cell Signaling Technology (1 mentions) Grp78, by Cell Signaling Technology (1 mentions) Thiazolyl blue tetrazolium bromide mtt, by Merck Group (1 mentions) Recombinant human vegf, by Merck Group (1 mentions) Bradford reagent, by Bio-Rad (1 mentions) Ripa buffer, by Cell Signaling Technology (1 mentions) Dimethyl sulfoxide (dmso), by Fujifilm (1 mentions) Phenylmethylsulfonyl fluoride pmsf, by Merck Group (1 mentions) Chemiluminescent substrate, by Cell Signaling Technology (1 mentions) 5 bromo 4 chloro 3 indolyl phosphate nitro blue tetrazolium bcip nbt, by Merck Group (1 mentions) Sodium fluoride naf, by Fujifilm (1 mentions) Bovine serum albumin bsa, by Fujifilm (1 mentions) Estradiol benzoate, by Merck Group (1 mentions) Coconut oil, by Merck Group (1 mentions) Coconut oil, by Research Diets (1 mentions)

Close spelling variants of this product

4-(2-aminoethyl) benzenesulfonyl fluoride hydrochloride (10 citations) 4-(2-Aminoethyl) benzenesulfonyl fluoride (AEBSF; (6 citations) 4-(2-aminoethyl) benzenesulfonyl fluoride (5 citations) Hydrochlorides (2 citations)

15 protocols using 4 2 aminoethyl benzenesulfonyl fluoride hydrochloride aebsf

1

Antibody Procurement and Cell Assays

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Antibodies against ATF6, CIP2A, GRP78, GAPDH, and β‐Actin were purchased from Cell Signaling Technology (Boston, MA, USA). Antibody against CIP2A for immunohistochemistry (IHC) was purchased from Novus Biologicals (Littleton, CO, USA), and for western blot analysis was purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Antibody against ERK was purchased from Zymed (Grand Island, NY, USA). Tunicamycin, cycloheximide, 3‐[4, 5‐dimethylthiazol‐ 2‐y1]‐2, 5‐diphenyltetrazolium bromide) (MTT), and 4‐(2‐Aminoethyl) benzenesulfonyl fluoride hydrochloride (AEBSF) were purchased from Sigma‐Aldrich (St. Louis, MO, USA).
Liu C., Hsu C., Huang T., Lee C., Chen J., Yang S., Jiang J., Chen W., Lee K, & Teng H. (2018). ER stress‐related ATF6 upregulates CIP2A and contributes to poor prognosis of colon cancer. Molecular Oncology, 12(10), 1706-1717.
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2

VEGF-induced Cell Proliferation Assay

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Recombinant human VEGF, thiazolyl blue tetrazolium bromide (MTT), 5-bromo-4-chloro-3-indolyl phosphate/nitro blue tetrazolium (BCIP/NBT), phenylmethylsulfonyl fluoride (PMSF), and 4-(2-Aminoethyl) benzenesulfonyl fluoride hydrochloride (AEBSF) were purchased from Sigma-Aldrich Corporation (St. Louis, MO, USA). RIPA buffer and chemiluminescent substrate were obtained from Cell Signaling Technology, Inc. (Beverly, MA, USA). Dimethyl sulfoxide, sodium fluoride (NaF), and bovine serum albumin (BSA) were from Wako Pure Chemical Industries (Osaka, Japan). Bradford reagent was purchased from Bio-Rad Laboratories, Inc. (Hercules, CA, USA). MRBH, also called MGN-3 or BioBran, was provided by Daiwa Pharmaceutical Co., Ltd. (Tokyo, Japan).
ZHU X., OKUBO A., IGARI N., NINOMIYA K, & EGASHIRA Y. (2016). Modified rice bran hemicellulose inhibits vascular endothelial growth factor-induced angiogenesis in vitro via VEGFR2 and its downstream signaling pathways. Bioscience of Microbiota, Food and Health, 36(2), 45-53.
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3

Estradiol Benzoate and High-Fat Diet Effects

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Estradiol benzoate (EB) and 4-(2-Aminoethyl) benzenesulfonyl fluoride hydrochloride (AEBSF, 1 mg/mL) were purchased from Sigma-Aldrich. Ketamine, Marcaine®, and Rimadyl® were purchased from Henry Schein Animal Health (Columbus, OH, USA). EB was dissolved in 100% ethanol (Sigma) prior to dissolving in coconut oil (Nature’s Way). Diets were purchased from Research Diets (New Brunswick, NJ): CON (10% kcal fat; D12450B) and 3 HFDs based on Research Diets 45% kcal fat diet D12451, with either 1%, 15% or 22.5% of kcals from LA, the remainder consisting of coconut oil SFAs. See Table 1 for FA profile of experimental diets.
Mamounis K.J., Hernandez M.R., Margolies N., Yasrebi A, & Roepke T.A. (2017). INTERACTION OF 17β-ESTRADIOL AND DIETARY FATTY ACIDS ON ENERGY AND GLUCOSE HOMEOSTASIS IN FEMALE MICE. Nutritional neuroscience, 21(10), 715-728.
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4

Intranasal Allergen Challenge in Mice

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Female BALB/c mice (Charles River, Margate, United Kingdom) and ST2−/− mice on a BALB/c background (a kind gift from Andrew McKenzie, MRC Laboratory of Molecular Biology, Cambridge, United Kingdom), 6 to 8 weeks old received 10 μg house dust mite (HDM) (Dermatophagoides pteronyssinus), Alternaria alternata, cat dander, or ragweed (Ambrosia artemisiifolia) extract (Greer, Lenoir, NC); papain or trypsin (Sigma-Aldrich, Dorset, United Kingdom); or 50 μL of vehicle, PBS intranasally. Mice were culled either 1 or 24 hours after challenge. In some experiments, the Alternaria extract was preincubated with either 4-(2-Aminoethyl)benzenesulfonyl fluoride hydrochloride (AEBSF) (25 mg/mL) or suramin (2 mM) (Sigma-Aldrich). FSSLYR-amide (Bachem AG, Bubendorf, Switzerland) was administered 30 minutes before allergen challenge (100 μg intraperitoneal and 12.5 μg intranasal). In the exacerbation protocol, mice were treated with either 15 μg HDM or PBS 3 times a week for 3 weeks before receiving single challenge with 10 μg Alternaria. All the experiments were performed in accordance with UK Home Office guidelines. Airway responsiveness was determined by direct measurements of lung function in anesthetized and tracheostomized mice 24 hours after final challenge.
Snelgrove R.J., Gregory L.G., Peiró T., Akthar S., Campbell G.A., Walker S.A, & Lloyd C.M. (2014). Alternaria-derived serine protease activity drives IL-33–mediated asthma exacerbations. The Journal of Allergy and Clinical Immunology, 134(3), 583-592.e6.
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5

Amyloid beta and Gut Hormone Detection

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Unfiltered nonpasteurized beer with a 9% alcohol content was purchased from Kukà S.r.L. (Italy). In addition, 95% v/v alcohol was purchased from Carsetti S.r.L. (Italy) and diluted to 9%. SafAleTM T-58 yeast containing Saccharomyces cerevisiae and emulsifier E491 was purchased from Fermentis (Italy). Protease inhibitors tosyl phenylalanyl chloromethyl ketone (TPCK) and 4-(2-aminoethyl) benzenesulfonyl fluoride hydrochloride (AEBSF or Pefabloc) were purchased from Sigma-Aldrich S.r.L. (Milano, Italy). The amyloid beta 40 mouse enzyme-linked immunosorbent assay (ELISA) kit and amyloid beta 42 mouse ELISA kit for Aβ(1-40) and Aβ(1-42) peptide determination in brain homogenates were purchased from Invitrogen (Camarillo, CA, USA). The Rat/Mouse Ghrelin (active) ELISA kit, Mouse Leptin ELISA, Rat/Mouse GIP (total) ELISA (Merk EZRMGIP-55K) and the multi-species GLP-1 Total ELISA (Merk EZGLP1T-36K) were bought from Merk group.
Cecarini V., Gogoi O., Bonfili L., Veneruso I., Pacinelli G., De Carlo S., Benvenuti F., D’Argenio V., Angeletti M., Cannella N, & Eleuteri A.M. (2022). Modulation of Gut Microbiota and Neuroprotective Effect of a Yeast-Enriched Beer. Nutrients, 14(12), 2380.
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6

Contractile Behavior of Human Airway Smooth Muscle

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4-(2-Aminoethyl)benzenesulfonyl fluoride hydrochloride (AEBSF), Rho-kinase inhibitor Y27632, MCh, and CCh were purchased from Sigma-Aldrich. Well characterized human ASM cells (14 (link)) were obtained through the gift of the Hope Organ and Tissue Donor Network. During growth and maintenance, the cells were cultured in serum-containing DMEM/F12 supplemented with 10% FBS (35–010-CV; Corning Life Sciences), 1% penicillin–streptomycin (P0781; Sigma-Aldrich), 1% l-glutamine (25030149; Thermo Fisher Scientific), 1% amphotericin B (15290018; Thermo Fisher Scientific), 0.17% 1 M CaCl2 × 2H2O, and 1.2% 1 M NaOH. Prior to force measurements, the cells were cultured under serum-deprived conditions by replacement of FBS with a 1% insulin–transferrin–selenium supplement (25–800-CR; Corning Life Sciences). All measurements were performed using cells at passage seven.
Redes J.L., Basu T., Ram-Mohan S., Ghosh C.C., Chan E.C., Sek A.C., Zhao M., Krishnan R., Rosenberg H.F, & Druey K.M. (2019). Aspergillus fumigatus–Secreted Alkaline Protease 1 Mediates Airways Hyperresponsiveness in Severe Asthma. ImmunoHorizons, 3(8), 368-377.
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7

Palmitate-induced ER stress signaling

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Dulbecco’s Modified Eagle’s Medium (DMEM), Ham’s nutrient mixture F-12, and fetal bovine serum (FBS) were purchased from Life Technologies (Gibco BRL, Grand Island, NY, USA). Palmitate and 4-(2-aminoethyl)benzenesulfonyl fluoride hydrochloride (AEBSF) were obtained from Sigma-Aldrich (St. Louis, MO, USA). The Palmitate was prepared as described previously [30 (link)]. Thapsigargin and GSK2606414 were purchased from Millipore (Billerica, MA, USA). p-IRE antibody (ab48187), ATF6 antibody (ab11909), PRMT1 antibody (ab3768), and PRMT4 antibody (ab110024) were purchased from Abcam (Cambridge, UK). caspase-3 antibody (#9662), p-eIF2α antibody (#9721), and CHOP antibody (#2895) were purchased from Cell Signaling Technology (Beverly, MA, USA). ASYM24 antibody (#07-414) was obtained from Millipore. β-actin antibody (sc-1616) and p-PERK antibody (sc-32577-R) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). All reagents were of the highest purity commercially available.
Park M.J., Han H.J, & Kim D.I. (2017). Lipotoxicity-Induced PRMT1 Exacerbates Mesangial Cell Apoptosis via Endoplasmic Reticulum Stress. International Journal of Molecular Sciences, 18(7), 1421.
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8

Metabolic Biomarkers in Fasted Plasma

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Blood samples were collected approximately 15 minutes after waking on the last day of each condition following an overnight fast; timing from the last meal remained constant across conditions. EDTA plasma was stored at −80C until the end of the study. 4-(2-Aminoethyl) benzenesulfonyl fluoride hydrochloride (AEBSF: Sigma Aldrich, St. Louis, MO) was added to the blood collection tubes to protect acylated ghrelin from degradation. All samples were run in duplicate in the same assay. Leptin was measured by radioimmunoassay (Millipore/Linco Research, Inc. St Charles, MO), which has a within assay coefficient of variation (CV) of 4.7% at a leptin concentration of 15.7 ng/mL. Acylated ghrelin was measured by double antibody sandwich ELISA (Millipore/Linco Research, Inc. St Charles, MO) which has a within assay CV of 3.2% at 397 pg/mL. Glucose was measured using the glucose oxidase method (Roche COBAS MIRA Clinical Analyzer) and insulin quantitated by radioimmunoassay (Millipore/Linco Research, Inc. St Charles, MO). HOMA-IR was calculated as fasting insulin (μU/ml) X fasting glucose (mg/dL) divided by 405, according to standard procedures.
Hart C.N., Carskadon M.A., Demos K.E., Van Reen E., Sharkey K.M., Raynor H.A., Considine R.V., Jones R.N, & Wing R.R. (2014). Acute changes in sleep duration on eating behaviors and appetite-regulating hormones in overweight/obese adults. Behavioral sleep medicine, 13(5), 424-436.
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9

Evaluating H. pylori Metabolic Hormones

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H. pylori-positive volunteers were instructed to arrive at the Endoscopic Unit of UMMC at 8am after 12 hours of fasting. Anthropometric measurements (height, weight, waist & hip circumference, and triceps-skinfold) were obtained. Validated Severity of Dyspepsia Assessment questionnaire (SODA) and SF-36 Health Survey were administrated to volunteers to assess baseline symptoms and quality of life profile.
Five ml of fasting blood was collected in EDTA-coated tube. The protease inhibitor, 4-(2-Aminoethyl) benzenesulfonyl fluoride hydrochloride (AEBSF) (Sigma-Aldrich, St. Louis MO), was added to the blood collection tubes at a concentration of 1 mg/ml. Tubes were kept on ice until centrifuged, and plasma was frozen at -80°C for metabolic hormone analysis.
Yap T.W., Leow A.H., Azmi A.N., Francois F., Perez-Perez G.I., Blaser M.J., Poh B.H., Loke M.F., Goh K.L, & Vadivelu J. (2015). Changes in Metabolic Hormones in Malaysian Young Adults following Helicobacter pylori Eradication. PLoS ONE, 10(8), e0135771.
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10

Serum Protease Inhibition Protocol

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A protease inhibitor solution contained 0.1 ml protease inhibitor cocktail (AEBSF, aprotinin, bestatin, EDTA, E-64, leupeptin; Sigma Aldrich), 0.1 g 4-(2-aminoethyl) benzenesulfonyl fluoride hydrochloride (AEBSF; Sigma Aldrich), and 0.9 ml dipeptidyl peptidase-4 (DDPIV) inhibitor solution (EMD Millipore Corporation, Billerica, MA). Blood collected at sacrifice was mixed 100:1 with the stock protease inhibitor. Blood cells and serum were separated by centrifugation (1,000g for 10 min) and serum was frozen at −70 °C. Serum specimens (100 μl) were examined using a Mouse Gut Hormone Panel (EMD Millipore Corporation) using a Luminex 200 analyser (Millipore). Two-sided Wilcoxon Rank-Sum tests were used for group comparisons.
Nobel Y.R., Cox L.M., Kirigin F.F., Bokulich N.A., Yamanishi S., Teitler I., Chung J., Sohn J., Barber C.M., Goldfarb D.S., Raju K., Abubucker S., Zhou Y., Ruiz V.E., Li H., Mitreva M., Alekseyenko A.V., Weinstock G.M., Sodergren E, & Blaser M.J. (2015). Metabolic and metagenomic outcomes from early-life pulsed antibiotic treatment. Nature Communications, 6, 7486.
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