To remove any leaf residue, a digestion solution comprising of 3.5% w/v sodium hydroxide (NaOH, S5881, Sigma-Aldrich) and 2.5% w/v sodium carbonate (Na2CO3, 222321, Sigma-Aldrich), in deionised water was used53 (link). The digestion solution was placed on a hotplate at 160 °C and stirred at low angular speed (80 to 120 rpm, depending on the size of the glassware and magnetic stirrer used) (SP88857105, Cimarec+, Thermo Scientific), until the sodium hydroxide and sodium carbonate were completely dissolved – approximately 15 minutes. The PDMS leaf imprint was placed into the digestion solution for 20 minutes. Once removed from the digestion solution, the PDMS leaf imprint was promptly rinsed thoroughly with deionised water. Any stubborn residue was carefully removed with tweezers with the PDMS leaf imprint submerged in deionised water (this was observed to occasionally around the edges of the PDMS leaf imprint). The PDMS leaf imprint was then placed in fresh deionised water for 20 minutes to remove any digestive solution residue. Finally, the imprint was rinsed thoroughly with deionised water and dried with dry nitrogen.
S5881
Manufactured by Merck Group
Sourced in Italy, United States, Germany
The S5881 is a laboratory instrument manufactured by Merck Group. It serves as a general-purpose device for various scientific applications. The core function of the S5881 is to perform measurements and analyses required in a laboratory setting. No further details or interpretations about its intended use are provided.
Automatically generated - may contain errors
Lab products found in correlation
6 protocols using s5881
1
PDMS Leaf Imprint Cleaning Protocol
2
Zoledronate Effects on PDLSC
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For all the experiments, PDLSCs were seeded at 4,000 cell/cm2 in MesenPro RS Medium added with ZOL (PHR1893, Sigma, Milan, Italy) dissolved in NaOH (S5881, Sigma, Milan, Italy) 0.1N. Cells cultivated in the absence of the drug were used as control. The ZOL concentration range chosen for this study was based on the literature and, above all, on the lack of information about the drug concentration in the alveolar bone. We tested the effects of ZOL between 0.1 and 100 μM. The observation of the maximum cytotoxic effect at 5 μM allowed us to set up 5 μM as the high concentration in our study.
3
Cycling Immunofluorescence Staining Protocol
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Cycling immunofluorescence was performed as described in (7 (link)). Briefly, cells were stained with dye-conjugated antibodies with nonoverlapping emission/excitation properties. A list of antibodies is provided in table S1. After overnight staining, cells were washed and imaged. To inactivate fluorescent dyes, the wells were bleached with 3% hydrogen peroxide (Sigma-Aldrich, 216763) and 20 mM sodium hydroxide (Sigma-Aldrich, S5881) in a base solution of phosphate-buffered saline. Plates were exposed to light for 1 hour. Plates were imaged after bleaching to confirm dye inactivation. In instances where dyes were not completely inactivated, another cycle of bleaching was performed. Plates were subsequently stained with additional antibodies. This process was repeated over several cycles.
4
Extracellular Matrix Solubilization Protocol
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dECM samples in
powder form were digested at designated concentrations (10, 15, and
20 mg/mL, w/v) in pepsin (Sigma, P6887) solution (1 mg/mL pepsin in
0.01 M HCl) (Merck, 100317) at room temperature under constant stirring
for 48 h. Upon completion, a part of all samples were spared as total
digests and the remaining samples were centrifuged at 5000g for 10 min. Supernatants were collected, labeled, and
used as soluble digests. Then, both total and soluble digests were
neutralized and buffered to physiological conditions (pH 7, 1X PBS)
by adding NaOH (Sigma, S5881) and 10X PBS. These pre-gel forms were
stored at −20 °C for further studies.
powder form were digested at designated concentrations (10, 15, and
20 mg/mL, w/v) in pepsin (Sigma, P6887) solution (1 mg/mL pepsin in
0.01 M HCl) (Merck, 100317) at room temperature under constant stirring
for 48 h. Upon completion, a part of all samples were spared as total
digests and the remaining samples were centrifuged at 5000g for 10 min. Supernatants were collected, labeled, and
used as soluble digests. Then, both total and soluble digests were
neutralized and buffered to physiological conditions (pH 7, 1X PBS)
by adding NaOH (Sigma, S5881) and 10X PBS. These pre-gel forms were
stored at −20 °C for further studies.
5
Quantifying Melanin in PIG1 Cells
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The melanin content of PIG1 cells was assessed following the previously established protocol (Zhou and Sakamoto 2020) (link). Briefly, PIG1 cells were seeded in a 6-well plate and harvested by trypsinization (trypsin, #T1300, Solarbio, Beijing, China). The supernatant was discarded after centrifugation at 1000 × g for 5 min. The cells were then lysed by adding 400 μL of NaOH solution (1 mol/L, #S5881, Sigma-Aldrich, MO, USA; containing 10% dimethyl sulfoxide) and heated in a metal bath at 70 °C for 1 h. Finally, the optical density (OD) was assessed at 490 nm on a microplate reader (DR-3518G, Hiwell Diatek, Wuxui, China).
6
Histological Evaluation of Decalcified Tendons
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After methanol fixation tendons were decalcified using 10 % EDTA solution (HNaO Sodium hydroxide S5881 Sigma, Germany; EDTA A3234 AppliChem, Germany) for three to six weeks under control by X-ray (30 kV and 10 s; X-ray equipment: Cabinet x-ray systems faxitron series, HP, Germany and X-ray film: Structurix D4DW 18 × 24, NDT Systems, Agfa, Germany). Then tendons were transferred to 100 % methanol. Twenty-four hours prior to cryosectioning, tissue samples were stored in a 5 % sucrose and phosphate-buffered saline solution. Sections were sliced to thickness of twelve micrometers (Cryostat CM 1950, Leica, Germany and Cryo Gel Embedding Medium, Leica, Germany). Hematoxylin and eosin staining was performed. On the basis of a semi-quantitative four-point-scoring system (from 0 = normal to 3 = markedly abnormal appearance) in six categories: (1) fibre arrangement, (2) tendon fibre structure, (3) regional variations in cellularity, (4) increased vascularity and (5) scarring (decreased collagen staining) and (6) hyalinisation were assessed as established by Longo et al [17 (link)]. The final score results from the sum of the individual scores given in each category.
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