DEAE-cellulose (10 g; Sigma Aldrich, Germany) dry gel washed with distilled water and left at 4°C for 16 h to remove small particles. The swollen gel was suspended in 0.5 M HCl (Merck Millipore, Germany) for 30 min and was filtered and washed with distilled water.The gel was suspended in 0.5 M NaOH (Merck Millipore, Germany) for 30 min (20 ) and washed with phosphate buffer (pH 6.3) 5×. The gel was packed into a XK 26/20 column (GE Healthcare, Sweden) with a 300 cm.h-1 linear velocity (26.5 mL.min-1) using preparative HPLC system (Waters, USA). The packed column was 85 mm bed height and 45 mL bed volume. The column was equilibrated by 3 column volume (CV), 0.07 M buffer phosphate pH 6.3 (19 ,21 (link)) at 225 cm.h-1 linear velocity (20 mL.min-1). Sample was loaded into the column at 150 cm.h-1. Linear flow rate of 13.3 mL.min-1 was applied to separate the adsorbed impurities.
0.5 m hcl
Manufactured by Merck Group
Sourced in United States
0.5 M HCl is a laboratory reagent that consists of a 0.5 molar solution of hydrochloric acid (HCl) in water. It is a commonly used solution for various applications in chemical and biological research, analysis, and experimentation.
Automatically generated - may contain errors
Lab products found in correlation
Kcl electrolyte solution, by Merck Group (2 mentions)
Surpass electrokinetic analyzer, by Anton Paar (2 mentions)
Deae cellulose, by Merck Group (1 mentions)
Xk26 20 column, by GE Healthcare (1 mentions)
0.5 m naoh, by Merck Group (1 mentions)
Preparative hplc system, by Waters Corporation (1 mentions)
0.1 m acetic acid, by Merck Group (1 mentions)
Nupage lds sample buffer 1x, by Thermo Fisher Scientific (1 mentions)
Precision plus protein dual color standard, by Bio-Rad (1 mentions)
7 protocols using 0.5 m hcl
1
DEAE-Cellulose Column Purification Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
DEAE-cellulose (10 g; Sigma Aldrich, Germany) dry gel washed with distilled water and left at 4°C for 16 h to remove small particles. The swollen gel was suspended in 0.5 M HCl (Merck Millipore, Germany) for 30 min and was filtered and washed with distilled water.The gel was suspended in 0.5 M NaOH (Merck Millipore, Germany) for 30 min (20 ) and washed with phosphate buffer (pH 6.3) 5×. The gel was packed into a XK 26/20 column (GE Healthcare, Sweden) with a 300 cm.h-1 linear velocity (26.5 mL.min-1) using preparative HPLC system (Waters, USA). The packed column was 85 mm bed height and 45 mL bed volume. The column was equilibrated by 3 column volume (CV), 0.07 M buffer phosphate pH 6.3 (19 ,21 (link)) at 225 cm.h-1 linear velocity (20 mL.min-1). Sample was loaded into the column at 150 cm.h-1. Linear flow rate of 13.3 mL.min-1 was applied to separate the adsorbed impurities.
+ Expand
DEAE-cellulose (10 g; Sigma Aldrich, Germany) dry gel washed with distilled water and left at 4°C for 16 h to remove small particles. The swollen gel was suspended in 0.5 M HCl (Merck Millipore, Germany) for 30 min and was filtered and washed with distilled water.The gel was suspended in 0.5 M NaOH (Merck Millipore, Germany) for 30 min (20 ) and washed with phosphate buffer (pH 6.3) 5×. The gel was packed into a XK 26/20 column (GE Healthcare, Sweden) with a 300 cm.h-1 linear velocity (26.5 mL.min-1) using preparative HPLC system (Waters, USA). The packed column was 85 mm bed height and 45 mL bed volume. The column was equilibrated by 3 column volume (CV), 0.07 M buffer phosphate pH 6.3 (19 ,21 (link)) at 225 cm.h-1 linear velocity (20 mL.min-1). Sample was loaded into the column at 150 cm.h-1. Linear flow rate of 13.3 mL.min-1 was applied to separate the adsorbed impurities.
2
Bovine Femoral Condyle Preparation for CMS
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To obtain the CMS, we collected the bovine femoral condyle, as previously reported (14 (link)). Briefly, the selected samples of 3 cm × 3 cm were carefully dissected and washed with water using anionic detergent. We then cut a triangular piece that was 1 cm on each side and 0.4–0.5 cm thick (Nukbone)®. Posteriorly, the biomaterial was demineralized with 0.5 M HCl (Merck, Millipore, United States) for 10 min and washed with distilled water to obtain the CMS (12 (link)), which was then sterilized using the hydrogen peroxide vapor/plasma sterilization method (20 (link)). The biomaterial was provided by the Materials Research Institute, Universidad Nacional Autónoma de México (UNAM).
3
Girdle Hydrogel Extraction in Chitons
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The scleritome and girdle were washed several times with distilled water to remove any possible dirt and particles. Shell plates were dissected from the body structure and the softened girdle structure was separated intact. Then to remove the calcareous shell layer on the surface of the girdle, specimens were treated with 0.5 M HCl (Sigma Aldrich, St. Louis, MO, USA) over 24 h. The integrity of the girdle structure was disrupted after the acid treatment, and pieces of girdle tissue were placed on dialysis membrane (Seamless Cellulose Tubing, size: 16/32, lot: 208001; Viskase Sales Corp., Chicago, IL, USA) and kept in distilled water with frequent changes of water over 3 days to ensure complete removal of the acid. Afterwards, the hydrogel samples were dried in an oven at 60 °C for 1 day. Although the same procedures were attempted, hydrogel formation was not observed in two other chiton species Plaxiphora aurata and Rhyssoplax olivacea.
4
Collagen Extraction and Electrophoresis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Between 1 and 2 g of the sample of interest were completely demineralized by overnight immersion in 0.5 M HCl (Sigma-Aldrich, Milano, Italy), dialyzed using Milli-Q water (Simplicity, Merck Millipore, Vimodrone, Italy) and lyophilized. Then, 50 mg were later solubilized in 1 mL 0.1 M acetic acid (Sigma-Aldrich) for 4 h at room temperature, using a thermoshaker set at 400 rpm (PHMT, Grant Instruments, Shepreth, UK). The extracted samples and a collagen standard (PureCol-S Bovine Collagen, Advanced Biomatrix, San Diego, CA, USA) were then diluted 1:10 using an electrophoresis buffer (NuPage LDS Sample Buffer 1X, Invitrogen, Rodano, Italy) containing a reducing agent (NuPage Reducing Agent 1X, Invitrogen). The solution was then kept for 10 min under shaking at 400 rpm and 70 °C using a thermoshaker (PHMT, Grant Instruments). Electrophoresis was carried out by loading an electrophoresis gel (NuPage Bis-Tris Mini Gels 10 Well, Invitrogen, ThermoFisher Scientific, Waltham, MA, USA) with the samples of interest together with a molecular weight marker (Precision Plus Protein Standard Dual Color, Bio-Rad, Segrate, Italy). The gel was run in a running buffer (MOPS SDS 1X, Invitrogen) using a 200 V electric field for 50 min. Staining followed according to a standard protocol using colloidal Coomassie Stains (Bio-Rad).
5
Zeta Potential Analysis of Membrane Surfaces
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Zeta potential is used to analyze the surface charge of membranes at different pH environments. It is particularly important to analyze the separation efficiency of membranes based on charge and also a confirmation test for surface modification [59 ]. Surface charge was analyzed by measuring the zeta potential using an Anton Paar SurPASS electrokinetic analyzer (Anton Paar, Ashland, VA, USA) in surface analysis mode. Before analysis, membranes were rinsed with copious amounts of DI water to remove any residual solvent or glycerol from the storage solution in the case of PBI membranes. The KCl electrolyte solution (Sigma Aldrich, St. Louis, MO) used in these measurements had an ionic strength of 1.0 mM. The pH values for the various readings were adjusted using 0.5 M HCl (Sigma Aldrich, St. Louis, MO, USA) and 0.5 M NaOH (Sigma Aldrich, St. Louis, MO, USA) solutions for acid and base titrations.
6
Zeta Potential Analysis of Membrane Surfaces
7
Quantifying Cellular Mineralization Potential
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The mineralization capability of cells was evaluated at day 14 of culture using Alizarin red staining and a quantitative calorimetric assay. For the staining, the samples were washed twice with phosphate-buffered saline and stained for 30 minutes using 1% Alizarin red (pH 6.3-6.4) at 37°C. The mineralized nodule area in red was measured using an image analyzer (ImageJ). For the quantification, cultures were washed with ddH 2 O and incubated overnight in 1 mL of 0.5M HCl (St. Louis, MO: Sigma-Aldrich) with gentle shaking. The solution was then mixed with o-cresolphthalein complexone in an alkaline medium (Stanbio LiquiColor; Boerne, TX: Stanbio), resulting in a purple calcium-cresolphthalein complexone complex that was quantified by absorbance measurement at a wavelength of 550-nm using a plate reader.
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!