P dimethylaminobenzaldehyde

Manufactured by Merck Group

Sourced in United States, Germany

P-dimethylaminobenzaldehyde is a chemical compound used as a laboratory reagent. It is a white to pale yellow crystalline solid. The compound is commonly used in various analytical and diagnostic procedures, particularly in the detection and quantification of certain classes of chemical compounds.

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Dimethylaminobenzaldehyde (11 citations) P-dimethylaminobenzaldehyde (PDAB) (2 citations) P-dimethylaminobenzaldehyde (DMAB) (2 citations)

46 protocols using p dimethylaminobenzaldehyde

Lipid-based Formulation Preparation and Characterization

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Phospolipon 90G (P90G) was purchased from Lipoid AG (Cologne, Germany) with the support of its Italian agent AVG srl. Cholesterol (CHOL), ESN, BRB and hydroxypropyl methylcellulose (HPMC) were provided by Sigma-Aldrich (Milan, Italy). Methanol (MeOH), methanol HPLC grade, acetonitrile (ACN), formic acid, dichloromethane (CH₂Cl₂), dimethylsulfoxide (DMSO) and formaldehyde solution, phosphate saline buffer (PBS), acetate buffer, NaOH, potassium borate, hyaluronidase from bovine testes Type IV-S, powder (mouse embryo tested, 750–3000 units/mg solid), compound 48/80 (condensation product of N-methyl-p-methoxyphenethylamine with formaldehyde, hyaluronic acid potassium salt from human umbilical cord, p-dimethylaminobenzaldehyde (98%, Ehrlich′s reagent) were purchased from Sigma-Aldrich (Milan, Italy). Piroxicam, progesteron, Prisma Buffer (P/N 110151), Hydration Solution (P/N: 120706) and Skin-PAMPA^TM system were purchased from pION Inc. (Billerica, MA, USA). Ultrapure water was produced by a synergy UV Simplicity water purification system provided by Merck KGaA (Molsheim, France). Phosphotungstic acid (PTA) was purchased from Electron Microscopy Sciences (Hatfield, PA, USA).

Vanti G., Bani D., Salvatici M.C., Bergonzi M.C, & Bilia A.R. (2019). Development and Percutaneous Permeation Study of Escinosomes, Escin-Based Nanovesicles Loaded with Berberine Chloride. Pharmaceutics, 11(12), 682.

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Antioxidant and Anti-inflammatory Assays

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Methanol, DPPH (2,2-diphenyl-1-picrylhydrazyl), ABTS (2,2′-Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)), TPTZ (2,4,6-tri(2-pyridyl)-s-triazine), ascorbic acid, hyaluronic acid (human umbilical cord), sodium hydroxide, hyaluronidase (bovine testes), calcium chloride, p-dimethylaminobenzaldehyde (PDMAB), and sodium borate were purchased from Sigma-Aldrich (USA). COX (ovine) and LOX inhibitor screening assay kits (Cayman Chemical Company, MI, USA), 96-well cell culture plate (Corning Life Sciences, Lowell, MA, USA), PMA (Sigma-Aldrich, St. Luis, MO, USA), and histamine release assay kit (SPI-Bio, France) were used.

Ashour R.M., El-Shiekh R.A., Sobeh M., Abdelfattah M.A., Abdel-Aziz M.M, & Okba M.M. (2023). Eucalyptus torquata L. flowers: a comprehensive study reporting their metabolites profiling and anti-gouty arthritis potential. Scientific Reports, 13, 18682.

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Colorimetric Assay for trans-4-Hydroxy-L-Proline

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An equal volume of 6 N HCl was added to samples or serial dilutions of trans-4-hydroxy-L-proline (ACROS Organics™, Morris Plains, NJ) and incubated overnight at 110°C.^{31 (link)} Samples were incubated for 5 min at room temperature following addition of 500 μL of oxidant solution (0.178 g Chloramines T in 15 mL of isopropanol and 10 mL of ddH₂O) and then 25 mL of acetate citrate buffer (120 g sodium acetate trihydrate, 12 mL acetic acid, 50 g citric acid monohydrate, and 34 g NaOH in 1 L ddH₂O, pH 6.0). Subsequently, 500 μL of Ehrlich's reagent (1 g p-dimethylaminobenzaldehyde (Sigma-Aldrich) in 20 mL isopropanol with 6.6 mL perchloric acid and 15.6 mL ddH₂O) was added and samples incubated at 60°C for 12 min. Following 4 min of cooling on ice, samples were transferred to a 96 well microtiter plate and absorbance read at 550 nm.

Duan W., Haque M., Kearney M.T, & Lopez M.J. (2017). Collagen and Hydroxyapatite Scaffolds Activate Distinct Osteogenesis Signaling Pathways in Adult Adipose-Derived Multipotent Stromal Cells. Tissue Engineering. Part C, Methods, 23(10), 592-603.

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Quantifying Type I Collagen Levels

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Hydroxyproline, which is directly proportional to type I collagen content, was measured by spectrophotometric determination8 ,9 (link). Briefly, the medium surrounding the gels was completely removed, and the gels were transferred to a glass tube (KIMAX; Fisher Scientific, St. Louis, MO, USA) with 2 mL of 6N HCl. O₂ was removed by ventilation with N₂ for 30 seconds. The gels were hydrolyzed at 110℃ for 12 hours. The samples were dried with a vacuum centrifuge and redissolved in distilled H₂O before measurement. Hydroxy-proline in the samples was reacted with oxidant (1.4% chloramines T in acetate-citric acid buffer; Sigma, St. Louis, MO, USA) and Ehrlich's reagent (0.4% p-dimethylaminobenzaldehyde; Sigma) in 60% perchloric acid (Fisher Chemical, Fair Lawn, NJ, USA) at 65℃ for 25 minutes, and hydroxyproline content was determined by spectrophotometer at 550 nm.

Kim S.H., Baek M.S., Yoon D.S., Park J.S., Yoon B.W., Oh B.S., Park J, & Kim H.J. (2014). Vitamin D Inhibits Expression and Activity of Matrix Metalloproteinase in Human Lung Fibroblasts (HFL-1) Cells. Tuberculosis and Respiratory Diseases, 77(2), 73-80.

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Scaffold Characterization Protocol

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On day 21, the scaffolds were harvested and measured for wet weight. They were then lyophilized and measured for dry weight. Swelling ratio was calculated as wet weight/dry weight of day 21 constructs. Each lyophilized sample was then digested in 500 μL of papainase solution (Worthington Biochemical) for 16 h at 60 °C. Once digested, the supernatant from each sample was used for the DNA, sGAG, and collagen content assays, as previously described.^{39 (link),40 (link)} DNA content was measured using the Quant-iT Picogreen dsDNA Assay Kit (Molecular Probes) and Lambda phage DNA as the standard. Fold proliferation was calculated as day 21 DNA content/day 1 DNA content. sGAG content was quantified using the 1,9-dimethylmethylene blue dye-binding assay with shark CS (Sigma-Aldrich) as the standard. Collagen content was measured using the Ehrlich’s reaction assay for hydroxyproline. Briefly, samples were acid hydrolyzed and reacted with p-dimethylaminobenzaldehyde and chloramine T (Sigma). Total collagen content was estimated assuming 1:7.46 hydroxyproline: collagen mass ratio.

Xu X.P, & Best P.M. (1990). Increase in T-type calcium current in atrial myocytes from adult rats with growth hormone-secreting tumors.. Proceedings of the National Academy of Sciences of the United States of America, 87(12), 4655-4659.

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Biochemical Analysis of Isoproterenol

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Isoproterenol hydrochloride, phytic acid, trichloroacetic acid (TCA), galactose, thiobarbituric acid (TBA), p-dimethyl amino benzaldehyde, and acetyl acetone reagent were purchased from Sigma Chemical Company, St. Louis, MO, USA. Glucose, uric acid, total protein and A/G ratio kits were purchased from Qualigens Diagnostics, Mumbai, India. All other chemicals used in this study were of analytical grade.

Brindha E, & Rajasekapandiyan M. (2015). Preventive Effect of Phytic Acid on Isoproterenol-Induced Cardiotoxicity in Wistar Rats. International Journal of Biomedical Science : IJBS, 11(1), 35-41.

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Collagen Content Quantification by Hydroxyproline Analysis

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Both the extracellular and intercellular collagen content were estimated according to Wöessner’s method. Cell cultures and media were dried in a laboratory oven. The total collagen level was evaluated based on the hydroxyproline level by hydrolysis of dried samples with 6N HCl at 100 ℃ for 24 h. Hydrolysates were neutralized by 5N NaOH. Samples of 0.2 mL were taken for further analysis and diluted with redistilled water to a final volume of 1 mL. Hydroxyproline was oxidized to pyrrole by 0.5 mL of chloramine T in a citrate buffer (pH 6.0), then shaken and incubated for 20 min at room temperature. In order to remove excess chloramine T, 0.5 mL of 3.15 m perchloric acid was added. After 5 min incubation at room temperature, the samples were treated with 0.5 mL of 20% p-dimethylaminobenzaldehyde (Sigma Aldrich, St. Louis, MO, USA) and incubated in 60 ℃ water bath for 20 min. The optical density was measured at 560 nm on a spectrophotometer

Gałdyszyńska M., Radwańska P., Szymański J, & Drobnik J. (2021). The Stiffness of Cardiac Fibroblast Substrates Exerts a Regulatory Influence on Collagen Metabolism via α2β1 Integrin, FAK and Src Kinases. Cells, 10(12), 3506.

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Quantification of Collagen via HYP Assay

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To determine the content of HYP, which is the marker for collagen, 2 to 3 microspheres were digested in a 100 μl digestion solution (pH 6.5) consisting of 50 mM phosphate buffer (Sigma), 5 mM EDTA (Sigma), 5 mM L-cysteine (Sigma), and 300 μg/ml papain (Sigma) at 60°C overnight, as described previously [9 (link)]. Then a digested sample was hydrolyzed with 6 N HCl at 110°C for 18 h in a hydrolysis tube (Pierre) after being flushed with nitrogen gas for 30 s and was neutralized by 6 N NaOH (pH 6~7). Neutralized samples were incubated with 50 μl of 0.05 M chloramine T solution (Sigma) for 20 min and oxidized by 50 μl of 3.15M perchloric acid (Sigma) for 5 min and finally mixed with 50 μl p-dimethylaminobenzaldehyde (20%, w/v; Sigma) for 20 min at 60°C for color development. The optical densities were measured at 557 nm with SaFire (TECAN) microplate reader. HYP content was estimated by linear interpolation using trans-4-hydroxy-L-proline (Sigma) as standard.

Han S., Li Y.Y, & Chan B.P. (2016). Extracellular Protease Inhibition Alters the Phenotype of Chondrogenically Differentiating Human Mesenchymal Stem Cells (MSCs) in 3D Collagen Microspheres. PLoS ONE, 11(1), e0146928.

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Colorimetric Assay for IDO1 Activity

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IDO1 activity was determined by colorimetric measure of the accumulated L-Kynurenine product in the cell culture medium. Cells were given L-Trp (2 mM) in phenol red free DMEM containing 10% FBS with or without δ-ALA (1 mM) and ferric citrate (100 μM). After 5 h of incubation the culture medium was de-proteinized by adding an equal volume of 3% trichloroacetic acid (Sigma # T6399) and incubation at 50 °C for 30 min. After centrifugation at 9000 × g for 10 min the supernatants were mixed with an equal volume of p-dimethyl-aminobenzaldehyde (Sigma # 109762; 20 mg/ml) in glacial acetic acid at room temperature and incubated 3 min to allow for chromophore formation. Absorbance at 492 nm was measured in a plate reader (Molecular Devices) and similarly-processed standards containing L-Kyn (Sigma #K8625) were used to calculate the sample L-Kyn concentrations.

Jayaram D.T., Sivaram P., Biswas P., Dai Y., Sweeny E.A, & Stuehr D.J. (2024). Heme allocation in eukaryotic cells relies on mitochondrial heme export through FLVCR1b to cytosolic GAPDH. Research Square.

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Immunohistochemical analysis of inflammatory markers

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Goat anti-rabbit horseradish peroxidase (HRP) (sc-2030; CA, USA), anti-TNF-α (sc-52746), anti-nitrotyrosine (sc-32757), anti-CD 45 (sc-1178), and 3,3′-diaminobenzidine (DAB) substrate (7304) were procured from Santa Cruz Biotechnology Inc. (TX, USA). Polyclonal anti-TGF-β1 (AV 37894), anti-iNOS (N7782), 2′,7′-dichlorofluorescein diacetate (DCF-DA), diaminofluorescein diacetate (DAF-2), dimethyl sulfoxide (DMSO), trichloroacetic acid (TCA), chloramine-T, sodium acetate, isopropanol, p-dimethylaminobenzaldehyde, perchloric acid, trans-4-hydroxy-l-proline standard, and all other required chemicals were obtained from Sigma Aldrich (St. Louis, MO, USA). Mayer’s hematoxylin and eosin (H&E) stain was purchased from Fisher Scientific (Pittsburgh, PA, USA).

Verma S., Kalita B., Bajaj S., Prakash H., Singh A.K, & Gupta M.L. (2017). A Combination of Podophyllotoxin and Rutin Alleviates Radiation-Induced Pneumonitis and Fibrosis through Modulation of Lung Inflammation in Mice. Frontiers in Immunology, 8, 658.

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