Intestine samples were homogenized in normal saline (Sinopharm Chemical Reagent Co., Ltd., Shanghai, China) at a ratio of 1 g sample: 9 mL normal saline. The homogenates were centrifuged at 10,000 × g for 20 min and the supernatant re-centrifuged at 10,000 × g for 30 min to obtain a clearer supernatant (at 4 °C temperature) which was analyzed for enzyme activities. Proteolytic activity was determined according to the Folin phenol reagent method of Kunitz (1947) (link) at 37 °C temperature and 7.5 pH. Lipase activity was determined at 37 °C according to procedures of the Nanjing Jiancheng Bioengineering Institute, test kit A054. Alpha-amylase activity was assayed according to the procedures of Bernfeld (1955) at 25 °C temperature and 6.9 pH. Proteolytic activity unit (U) was expressed as 1 μg tyrosine released per minute; lipase activity unit (U) as 1 μmol substrate consumed per minute; and amylase activity unit (U) as 1 μmol of maltose released per minute. Protein concentration in supernatant was determined with the Coomassie brilliant blue method of Bradford (1976) (link).
Normal saline
Manufactured by Sinopharm
Sourced in China
Normal saline is a sterile, isotonic solution composed of sodium chloride (0.9% w/v) in water. It is commonly used as a general-purpose fluid and electrolyte replenishment solution in various medical and laboratory settings.
Automatically generated - may contain errors
Lab products found in correlation
Triton x 100, by Sinopharm (1 mentions)
Sodium chloride (nacl), by Sinopharm (1 mentions)
Phosphate buffered saline (pbs), by Sinopharm (1 mentions)
Tryptone, by Sinopharm (1 mentions)
Acetic acid, by Sinopharm (1 mentions)
Calcium sulfate, by Sinopharm (1 mentions)
Glycerin, by Sinopharm (1 mentions)
Maltose, by Sinopharm (1 mentions)
Sorbitol, by Sinopharm (1 mentions)
Kanamycin, by Sinopharm (1 mentions)
Ampicillin, by Sinopharm (1 mentions)
Yeast extract, by Sinopharm (1 mentions)
K2hpo4 3h2o, by Sinopharm (1 mentions)
Mgso4 7h2o, by Sinopharm (1 mentions)
Sodium acetate, by Sinopharm (1 mentions)
Trisodium citrate, by Sinopharm (1 mentions)
Mgcl2, by Sinopharm (1 mentions)
Trichloroacetic acid, by Sinopharm (1 mentions)
Peptone, by Sinopharm (1 mentions)
Tris buffer, by Sangon (1 mentions)
3 protocols using normal saline
1
Intestinal Enzyme Activity Assay
2
Diverse Biological Reagents Acquisition
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Neutral protease, papain, chitosan, bovine insulin, myoglobin, and tryptophan were purchased from Aladdin Reagent Co., Ltd. (Shanghai, China). Tryptone, yeast extract, Agar, NaCl, glycerin, K2HPO4·3H2O (AR), sorbitol, MgSO4·7H2O (AR), trisodium citrate, MgCl2 (AR), kanamycin, ampicillin, peptone, maltose, sodium acetate, calcium sulfate, acetic acid, Triton X-100, phosphate-buffered saline, normal saline, trichloroacetic acid, and other reagents (AR) were purchased from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China).
3
DF Compound Extraction and Characterization
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Chloral hydrate, sodium chloride injection, normal saline, and methanol were bought from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China). Glycine, TRIS buffer, and sodium dodecyl sulfate (SDS) was purchased from Sangon Biotech (Shanghai) Co., Ltd. (Shanghai, China). Skim milk powder was purchased from BBI Life Sciences (Shanghai, China). 30% acrylamide and ammonium persulfate (AP) were bought from Beijing Dingguo Changsheng Biotechnology Co., Ltd. (Beijing, China). Tetramethylethylenediamine (TEMED) was provided by Sigma (St. Louis, MO, United States). GeneView (Ste. Genevieve, MO, United States) provided the enhanced chemiluminescence (ECL) Kit. The Milli-Q (18.2 MΩ) ultra-pure water system (Millipore, Billerica, MA, United States) was used to prepare pure water. Mouse serum interferon gamma (IFN-γ), IL-10, IL-12p70, IL-17A, TNF-α, IL-6, monocyte chemoattractant protein-1 (MCP1), and IL-1β enzyme-linked immunosorbent assay (ELISA) kits were purchased from Bender (Gruenberg, Germany).
DF was prepared in our laboratory [Batch No.: 20180522. The DF had a rhodojaponin III content of 41.6% and a rhodojaponin VI content of 13.6%, as determined by the linear curve method (Yao et al., 2019 )]. Dexamethasone was purchased from GlpBio (Batch No.: GC40775. Montclair, CA, United States).
DF was prepared in our laboratory [Batch No.: 20180522. The DF had a rhodojaponin III content of 41.6% and a rhodojaponin VI content of 13.6%, as determined by the linear curve method (Yao et al., 2019 )]. Dexamethasone was purchased from GlpBio (Batch No.: GC40775. Montclair, CA, United States).
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