The assay employed was based on methods previously described in literature. 8 In brief, human skin model tissue was homogenized and solubilized in 0.1% Triton-X 100, 0.2 M Tris-HCl (pH 8.0) buffer, followed by ultrasonication and subsequent centrifugation at 2000 x g for 10 minutes to obtain supernatants for enzyme assay. To measure elastase activity, 2 μL of substrate (62.5mM, N-succinyl-tri-alanyl-p nitroanilide from Sigma-Aldrich) was incubated with 100 μL lysate solution at 37°C for 2 hours. The amount of released nitroaniline was measured by determining absorbance at 410 nm using a spectrophotometer (Molecular Devices).
N succinyl tri alanyl p nitroanilide
Manufactured by Merck Group
Sourced in United States
N-succinyl-tri-alanyl-p-nitroanilide is a chemical compound used as a substrate in laboratory settings. It is a synthetic peptide that can be utilized to measure the activity of certain enzymes.
Automatically generated - may contain errors
Lab products found in correlation
Dulbecco s minimal essential medium dmem, by Thermo Fisher Scientific (1 mentions)
Acrylamide and n n methylenebisacrylamide, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Sodium dodecyl sulfate (sds), by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Trypsin, by Thermo Fisher Scientific (1 mentions)
3 protocols using n succinyl tri alanyl p nitroanilide
1
Elastase Activity Measurement in Skin
2
Cell Viability Assay Using MTT
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RA was a product of BIOKOM (Warsaw, Poland). BP-3, radioimmunoprecipitation assay (RIPA) buffer, [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] (MTT), dimethyl sulfoxide (DMSO), sodium dodecyl sulfate (SDS), bovine serum albumin (BSA), gelatin, acrylamide and N,N’-Methylenebisacrylamide, protease inhibitor cocktail (P8340), and N-succinyl-tri-alanyl-p-nitroanilide (STANA) were provided by Sigma-Aldrich Corp. (St. Louis, MO, USA). Dulbecco’s minimal essential medium (DMEM), phosphate-buffered saline (PBS), and fetal bovine serum (FBS) used in the cell culture were purchased from Gibco (Thermo Fisher Scientific, Waltham, MA, USA). Penicillin, streptomycin, and glutamine were obtained from Quality Biologicals Inc. (Gaithersburg, MD, USA).
3
Peptide Synthesis Protocol with DMEM
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Dulbecco's Modified Eagle's Medium (DMEM), penicillin/streptomycin, fetal bovine serum (FBS), phosphate-buffered saline (PBS), and trypsin were purchased from Gibco (Carlsbad, CA). Hydroxybenzotriazole (HOBt), diisoproylcarbodiimide (DIC), 1,8-diazabicyclo[5.4.0]undec-7-en (DBU), N,N-diisopropylethylamine (DIEA), gallic acid and N-succinyl-tri-alanyl-p-nitroanilide were obtained from Sigma-Aldrich (St. Louis, MO). l -Form amino acids (Fmoc-Lys (Boc)–OH, Fmoc-Thr (tBu)–OH, and Fmoc-Pro–OH) were purchased from Bead-Tech (Seoul, Korea).
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