The proline concentration was determined according to Ábrahám [64 (link)] with slight modification. Fifty mg of dry homogenized Artemisia leaves were homogenized with 10 mL of 3% sulfur salicylic acid in a mortar. The mixture was filtered and 1 mL of the resulting filtrate, 2 mL of ninhydrin reagent, and 2 mL of acetic acid were heated at 95 °C for 1 h. The proline concentration was assessed using the absorption value of the reaction mixture at 505 nm and a calibration curve with 5 different proline (Merck) concentrations.
Proline
Manufactured by Merck Group
Sourced in United States, Germany, United Kingdom, Japan, China, Belgium, Switzerland, Spain, Australia, Italy
Proline is a lab equipment product manufactured by Merck Group. It is a versatile instrument designed for performing various laboratory tasks. The core function of Proline is to provide accurate and reliable measurements and analysis in a wide range of scientific applications.
Automatically generated - may contain errors
Lab products found in correlation
Dexamethasone (dex), by Merck Group (63 mentions)
Alanine, by Merck Group (32 mentions)
Phenylalanine, by Merck Group (31 mentions)
Valine, by Merck Group (31 mentions)
Ascorbic acid, by Merck Group (31 mentions)
Tyrosine, by Merck Group (28 mentions)
Serine, by Merck Group (28 mentions)
Ascorbate 2 phosphate, by Merck Group (28 mentions)
Threonine, by Merck Group (27 mentions)
Isoleucine, by Merck Group (27 mentions)
Leucine, by Merck Group (27 mentions)
Glutamic acid, by Merck Group (26 mentions)
Glycine, by Merck Group (25 mentions)
Aspartic acid, by Merck Group (24 mentions)
Histidine, by Merck Group (24 mentions)
Methionine, by Merck Group (23 mentions)
Sodium pyruvate, by Merck Group (22 mentions)
Sodium pyruvate, by Thermo Fisher Scientific (20 mentions)
Tryptophan, by Merck Group (19 mentions)
Glutamine, by Merck Group (16 mentions)
238 protocols using proline
1
Proline Determination in Artemisia Leaves
2
Hypertrophic Differentiation of Human MSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A high-density pellet culture system was applied for the chondrogenic differentiation of human MSCs, as described previously.54 (link) Cells were induced for chondrogenic predifferentiation for 14 d in pellet culture (200 000 cells/pellet) in chondrogenic medium consisting of high-glucose DMEM with 50 μg·mL−1 ascorbate acid 2-phosphate (Sigma-Aldrich), 40 μg·mL−1 proline (Sigma-Aldrich), 100 nmol·L−1 dexamethasone (Sigma-Aldrich), 10 ng·mL−1 recombinant human TGFβ3 (R&D systems), and 1% ITS Universal Culture Supplement Premix (BD Biosciences, San Jose, CA, USA).
To further induce hypertrophic differentiation, chondrogenic differentiated pellets were exposed to hypertrophic differentiation medium consisting of high-glucose DMEM with 50 μg·mL−1 ascorbate acid 2-phosphate, 40 μg·mL−1 proline, 1 nmol·L−1 dexamethasone, 1% ITS Universal Culture Supplement Premix, and 1 nmol·L−1 triiodothyronine (Sigma-Aldrich) for 14 d as described previously.52 (link) The medium was changed every 3 d.
To further induce hypertrophic differentiation, chondrogenic differentiated pellets were exposed to hypertrophic differentiation medium consisting of high-glucose DMEM with 50 μg·mL−1 ascorbate acid 2-phosphate, 40 μg·mL−1 proline, 1 nmol·L−1 dexamethasone, 1% ITS Universal Culture Supplement Premix, and 1 nmol·L−1 triiodothyronine (Sigma-Aldrich) for 14 d as described previously.52 (link) The medium was changed every 3 d.
3
Chondrogenic and Hypertrophic Differentiation of BMSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For chondrogenic and hypertrophic differentiation,14 (link) 1 × 106 BMSCs were placed in polypropylene tubes, centrifuge at a speed of 500 g for 20 min. Then remove the medium and add 500 μL of low-glucose DMEM supplemented with 10−7 M dexamethasone (Sigma–Aldrich), 1%(v/v) ITS (Sigma–Aldrich), 50 μM ascorbate-2-phosphate (Sigma–Aldrich), 1 mM sodium pyruvate, 50 μg/ml proline (Sigma–Aldrich) and 20 ng/ml TGFβ3. Change the medium three times a week and culture for 14d. Then, the serum-free chondrogenic medium was replaced with hypertrophic medium supplemented with 50 nM thyroxine (Sigma–Aldrich), 10−8 M dexamethasone, 250 μM ascorbate-2-phosphate, 1 mM sodium pyruvate and 50 μg/ml proline. Change the medium three times a week and culture for another 14d.
4
Proline Analysis in Leaf Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
proline analysis was performed according to the methods outlined by Bates, Waldren & Teare (1973) (link). The leaves were collected before the drying treatment every five days for 30 days (seven times in total) after watering ceased. After collecting 0.1 g (15 total repetitions) of each leaf, 10 mL of a sulfosalicylic acid solution (3%, w/v) was added, followed by mortar grinding. The grinding solution was filtered with two layers of filter paper (Whatman No. 42). After adding 1 mL of glacial acetic acid and 1 mL of ninhydrin reagent to 1 mL of the filtrate, the test tube was capped, reacted in boiling water (100 °C) for one hour, and then stored at room temperature (21.0 °C) for five minutes. Then, two mL of toluene was added, stirred for 20 s, and then the supernatant was taken, and the wavelength was measured at 520 nm using a UV spectrophotometer (X-ma 2000, Human Crop.). Quantitation was calculated according to a calibration curve prepared using proline (Sigma-Aldrich Co., St. Louis, MO, USA) as the standard material and expressed as µmol proline/g FW.
5
Proline Quantification in Leaf Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Proline accumulation in leaf tissue was determined via reaction with ninhydrin (Sigma Aldrich), as described by Bates et al. [79 (link)]. Purified Proline (Sigma Aldrich, Melbourne, VIC, Australia) was used to build a standard curve for Proline content quantification. Approximately 0.5 fresh leaf samples were homogenized in 10 mL of 3% aqueous sulfosalicylic acid and centrifuged at 3000 rpm for 1 min. Exactly 2 mL of supernatant was reacted with 2 mL of ninhydrin acid and 2 mL of glacial acetic acid for 1 h at 100 °C in a heater. The chromophore was extracted using 2 mL of Toluene, and its absorbance at 520 nm was determined by Genesys 10-s UV/Vis Spectrophotometer (Thermo Spectronic, Waltham, MA, USA) with toluene used as blank. Proline content was calculated using the following formula.
6
Mesenchymal Stem Cell Chondrogenesis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Example 7
ASC Cell Culture
Once first passaged ASCs reached 90% confluence, ASC monolayers and microbeads were then treated for 5 days with MSCGM, MSCGM with 10−7 M 24R,25-dihydroxyvitamin D3, chondrogenic medium (CM) consistent of high glucose DMEM with 1 mM sodium pyruvate (Mediatech, Manassas, Va., USA), 40 μg/ml proline (Sigma), 50 μg/ml ascorbate-2-phosphate (Sigma), 1% ITS+, 100 nM dexamethasone (Sigma), 10 ng/ml recombinant human transforming growth factor beta-I (TGF-31, R&D Systems, Minneapolis, Minn., USA) and 100 ng/ml recombinant human bone morphogenic protein 6 (BMP-6, PeproTech, Rocky Hill, N.J., USA) or medium consistent of high glucose DMEM with 1 mM sodium pyruvate (Mediatech, Manassas, Va., USA), 40 μg/ml proline (Sigma), 50 μg/ml ascorbate-2-phosphate (Sigma), 1% ITS+, and 10−7M 24R,25-dihydroxyvitamin D3. Once media were changed on the fifth day, RNA was collected after 8 hours as described below while media and ASCs lysed in 0.05% Triton X-100 were collected after 24 hours. Monolayer fourth passaged chondrocytes cultured in DMEM, 10% FBS, and 50 μg/mL ascorbic acid and Sprague Dawley-derived clone 9 liver cells (ATCC, Manassas, Va., USA) cultured in F12K medium and 10% FBS served as controls. All media contained 1% penicillin and streptomycin.
7
Proline Content Quantification in Leaves
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Proline content was detected in fresh leaves after 40 DAS according to the method described by Naidu et al. [49] . Briefly, 0.5 g of the leaves were transferred to tubes containing 5 mL of a methanol:chloroform:distilled water (60:25:15) mixture. The tubes were heated at 60 • C for 2 min and the mixtures were centrifuged at 10,000 rpm for 10 min. To the supernatant (1 mL) was added 4 mL of ninhydrin solution, 4 mL of glacial acetic acid, and 1 mL of distilled water. Then, the mixture was heated at 90 • C for 45 min, cooled to room temperature and absorbance was determined at 520 nm (SPEKOL 1300 UV VIS spectrophotometer, Analytik Jena). Proline (Sigma) was used as reference standard (y = 0.0314x + 0.0409; R 2 = 0.9993) and the results were expressed as µg Proline g fresh weight -1 (µg Pro g FW -1 ).
8
Proline Content Quantification in Plants
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The proline contents of the leaf and roots of the sample seedlings were determined according to the method of Claussen [9] . The absorbance of the mixture at 546 nm were obtained with a spectrophotometer. The proline concentration was determined from a calibration curve obtained with proline (Sigma) and calculated as a fresh weight basis [μmol proline (g FW -1 )].
9
Chondrogenic Differentiation of Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A 250 μL volume of cell suspension (containing 2.5 × 105 cells, including both living and dead cells) was added to six 15 mL tubes (Falcon) containing a chondrogenic induction medium consisting of Dulbecco's Modified Eagle Medium (DMEM; Thermo Fisher Scientific), 10 ng/mL transforming growth factor-β3 (TGF-β3, Miltenyi Biotec, Bergisch Gladbach, Germany), 500 ng/mL bone morphogenetic protein 2 (BMP-2, Medtronic, Minneapolis, MN, USA), 40 μg/mL proline (Merck), 100 nM dexamethasone, 100 μg/mL pyruvate (Merck), 1% antibiotic–antimycotic, 50 μg/mL ascorbate-2-phosphate, and 1% ITS + Premix (Becton Dickinson, San Jose, CA, USA). The cells were centrifuged at 450×g for 10 min to form cell pellets, which were cultured for 21 days. The cultured cell pellets were photographed and weighed with a semi micro balance (CPA225D, Sartorius, Gottingen, Germany). The pellets were cut into 5 µm sections and stained with safranin O and toluidine blue.
10
Metal-Ligand Complexation Synthesis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Pefloxacin mesylate dihydrate (HPf), 1,10-phenanthroline (phen), and 2,2′-bipyridine (bipy) were purchased from Sigma–Aldrich Chemical Co. Alanine, proline, aspartic acid, copper nitrate trihydrate, potassium hydroxide, potassium dihydrogen orthophosphate, and sodium hydroxide were used as received from Merck Chemical Company. Nitric acid (68%), Hydrochloric acid, dimethyl sulfoxide (DMSO, Aldrich), acetone, diethyl ether, and ethanol (BDH-PROLABO), were of analytical grade.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!