Benzoyl arginine p nitroanilide

Manufactured by Merck Group

Sourced in United States

Benzoyl arginine p-nitroanilide is a synthetic substrate used in biochemical assays and research applications. It is a chromogenic compound that undergoes enzymatic cleavage, resulting in the release of a colored product that can be measured spectrophotometrically. This substrate is commonly used to detect and quantify the presence of certain proteolytic enzymes, such as trypsin and other serine proteases.

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Lab products found in correlation

Advia 1650 chemistry system, by Siemens (1 mentions) Cysteine hcl, by Merck Group (1 mentions) Hepes, by Biowest (1 mentions)

Spelling variants of this product

Nα-benzoyl-DL-arginine-p-nitroanilide (7 citations) N-α-benzoyl-DL-arginine-p-nitroanilide (BApNA), (4 citations) Nα-benzoyl-DL-arginine p-nitroanilide hydrochloride (BApNA; (3 citations) Nα-benzoyl-DL-arginine-p-nitroanilide hydrochloride (3 citations) Benzoyl-arginine-p-nitroanilide (BApNA; (2 citations)

3 protocols using benzoyl arginine p nitroanilide

Gastrointestinal Enzyme and Bile Analysis

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Trypsin activity and total bile salt levels were measured in pooled freeze-dried gastrointestinal contents from the four separate intestine segments (IN1–4). Trypsin activity was determined colorimetrically as described by Kakade and co-workers^{(26 (link))} using the substrate benzoyl arginine p-nitroanilide (Sigma no. B-4875, Sigma Chemical Co., St. Louis, MO) and a curve derived from standardised bovine trypsin solution.
Bile salt levels were determined using the enzyme cycling amplification/Thio – NAD method (Inverness Medical, Cheshire, UK) in the ADVIA®1650 Chemistry System (Siemens Healthcare Diagnostics Inc.) at the Central Laboratory of the Faculty of Veterinary Medicine, Norwegian University of Life Sciences, Oslo.

Zhou W., Lie K.K., Chikwati E., Kousoulaki K., Lein I., Sæle Ø., Krogdahl Å, & Kortner T.M. (2023). Soya saponins and prebiotics alter intestinal functions in Ballan wrasse (Labrus bergylta). The British Journal of Nutrition, 130(5), 765-782.

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Quantifying Arg-Gingipain Activity in P. gingivalis

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Gingipains are cysteine proteinases secreted by P. gingivalis, which are related to the virulence of the bacteria since they play a critical role in tissue destruction. To determine the Arg-gingipain proteolytic activity, 10 μL of bacterial suspension with the different treatments, after 10 h of incubation, were mixed with 50 μL of assay buffer (10 mM cysteine- Hydrochloric acid (HCL), 1 M HEPES (Biowest), pH 7.5) in microtiter plates. To each well, 100 μL of substrate solution (0.5 mM benzoyl-arginine p-nitroanilide (Sigma-Aldrich, St. Louis, MO, USA), 10 mM cysteine-HCL, 50 mM Tris-HCl, pH 7.5) were added. After incubation for 16 h at 37 °C, the absorption at 405 nm was measured (PowerWave Ht). Three independent experiments were performed, with two replicates at each condition (n = 6).

Munar-Bestard M., Llopis-Grimalt M.A., Ramis J.M, & Monjo M. (2021). Comparative In Vitro Evaluation of Commercial Periodontal Gels on Antibacterial, Biocompatibility and Wound Healing Ability. Pharmaceutics, 13(9), 1502.

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Analysis of Digestive Enzyme Activities

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Trypsin activity was analyzed according to the method of Kakade et al. (1973) (link) using benzoyl arginine p-nitroanilide (No. B-4875, Sigma Chemical Co., St. Louis, MO) as the substrate. Total bile acid levels were determined at the Central Laboratory of Faculty of Veterinary Medicine of NMBU according to the method of Krogdahl et al. (2020) (link).
The capacity of brush border leucine aminopeptidase (LAP) was analyzed as the description of Bieth et al. (Bieth et al., 1974) (link). The tissue homogenate was prepared using ice-cold Tris -mannitol buffer (1:20, w/v). The four-(two-aminoethyl)benzene-sulfonyl fluoride hydrochloride (Pefabloc SC, Basel, Switzerland) was used as a serine proteinase inhibitor to prevent protein hydrolysis during sample preparation. The incubations were performed at 37°C according to the method of Krogdahl et al., 2003 (link). The LAP capacity is expressed as mmol substrate hydrolyzed per intestinal section per hour per kg fish.

Wang J., Jaramillo-Torres A., Li Y., Brevik Ø.J., Jakobsen J.V., Kortner T.M., & Krogdahl Å. (2022). Gut Health and Microbiota in Out-of-Season Atlantic Salmon (Salmo salar L.) Smolts Before and After Seawater Transfer Under Commercial Arctic Conditions: Modulation by Functional Feed Ingredients. Frontiers in Marine Science, 9.

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