Porcine skin

Manufactured by Merck Group

Sourced in United States

Porcine skin is a lab equipment product used for various research and testing purposes. It serves as a substrate for various studies and experiments. The core function of porcine skin is to provide a standardized and readily available material for these applications.

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Gelatin from porcine skin (105 citations) Type A gelatin from porcine skin (80 citations) Porcine skin gelatin (46 citations) Type A porcine skin gelatin (30 citations) Porcine skin gelatine (8 citations) Gelatine from porcine skin (7 citations) Gelatin (GEL) from porcine skin (2 citations) Type A from porcine skin (2 citations) Porcine skin gelatin solution (2 citations) Porcine skin-derived gelatin (2 citations)

7 protocols using porcine skin

Methacrylated Gelatin Synthesis

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Methacrylated groups were grafted to gelatin as previously reported [33 (link)]. Briefly, gelatin derived from porcine skin (Sigma-Aldrich) was dissolved at 10% (w/v) in PBS at 60 °C. After obtaining a clear solution, 0.14 ml of methacrylic anhydride (Sigma-Aldrich) for each gram of gelatin was added dropwise and the reaction was carried on for 3 h at 50 °C. The GelMA solution was then dialyzed against deionized water using 12–14 kDa cutoff dialysis tubes (VWR scientific) for 6 days at 50 °C to remove unreacted methacrylic anhydride and any additional by-products. GelMA was lyophilized and stored at −20 °C until further use.

Tognato R., Parolini R., Jahangir S., Ma J., Florczak S., Richards R.G., Levato R., Alini M, & Serra T. (2023). Sound-based assembly of three-dimensional cellularized and acellularized constructs. Materials Today Bio, 22, 100775.

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Immortalized Lung Adenocarcinoma Cell Culture

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For cell studies, we used an immortalized human lung adenocarcinoma epithelial cell line A549 (Department of Medical Genetics, School of Basic Medical Sciences, Jilin University). Dulbecco’s modified Eagle’s medium (DMEM), penicillin-streptomycin, trypsin, and fetal bovine serum were purchased from Sigma (USA). Cell Counting Kit-8 (CCK-8; Sigma, USA), acetone, anhydrous alcohol, GEN, ICA, gelatin from porcine skin (gel strength 250 g Bloom, Type A), glutaraldehyde, and acetone were purchased form Sigma (USA). A Zorbax® SB-C₁₈ high-performance liquid chromatography (HPLC) column (5 μm; 4.6 × 150 mm) for FLA analysis was purchased from Agilent Technologies (USA). HPLC-grade solvents methanol and acetonitrile were purchased from Sigma-Aldrich and Fisher Scientific (USA). All other solvents and chemicals used were in high-grade purity. Fluorescence inverted microscope (Olympus, CKX41, Japan), desktop centrifuge (Eppendorf, Mini Spin, German), JB-2 constant temperature magnetic stirrer (Shanghai Lei Xin Instrument Co., Ltd., China), KQ-100DA ultrasonic oscillators (Kunshan Ultrasonic Instrument Co., Ltd. China), transmission electron microscope (TEM; JEM-1011, Japan), SEM (Hitachi S-4800, Japan), and AFM (VEECO Dimension icon, USA) were used.

Song X., Gan K., Qin S., Chen L., Liu X., Chen T, & Liu H. (2019). Preparation and characterization of general-purpose gelatin-based co-loading flavonoids nano-core structure. Scientific Reports, 9, 6365.

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Gelatin Methacrylate Synthesis Protocol

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Gelatin methacrylate was synthesized according to an earlier protocol.^{[56 (link)]} Briefly, a 10% w/v gelatin (porcine skin, Sigma) solution in phosphate buffered saline (PBS) was prepared and heated to 60 °C with constant mixing. After the gelatin was dissolved completely, the temperature was reduced to 50 °C and allowed to reach steady state. After the solution temperature reached 50 °C, methacrylic anhydride (Sigma) was slowly added to the solution to achieve a 5:1 volumetric ratio of gelatin solution : methacrylic anhydride solution. The typical basis reaction volume consisted of 50 ml gelatin solution. Subsequently, the mixture was allowed to react for one hour at 50 °C with continual mixing. After one hour, warmed PBS (40 °C) heated in a secondary beaker was added at a 4:1 volume ratio to the gelatin-methacrylic anhydride solution to deactivate the reaction. Subsequently, the resulting mixture was added to 10 kDa dialysis tubing and the tubing was placed in reverse osmosis water and allowed to dialyze for one week. To ensure effective separation, the dialysis solution was replaced with fresh solution daily. Following the dialysis procedure, the gelatin mixture was lyophilized until dry.

Johnson B.N., Lancaster K.Z., Zhen G., He J., Gupta M.K., Kong Y.L., Engel E.A., Krick K.D., Ju A., Meng F., Enquist L.W., Jia X, & McAlpine M.C. (2015). 3D Printed Anatomical Nerve Regeneration Pathways. Advanced functional materials, 25(39), 6205-6217.

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Fabrication of HA/Gelatin Composites

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Nanosized hydroxyapatite powder (<200 nm particle size, nGimat) and type-A gelatin (porcine skin, Sigma Aldrich) were used to fabricate HA/gelatin composites. Gelatin powder was dissolved in distilled water in a rotational shaker at 50 °C and mixed with 4.5, 7, or 12 g of HA powder (Table 1). The mixture was placed in a planetary centrifugal mixer (AR-100, Thinky) for 1–2 min to produce homogeneous HA/gelatin composites.

Lee H., Yoo J.M, & Nam S.Y. (2021). Additive Fabrication and Characterization of Biomimetic Composite Bone Scaffolds with High Hydroxyapatite Content. Gels, 7(3), 100.

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Gelatin Degradation by MMP-14

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10 nM cdMMP-14 was incubated with 1 mg/mL gelatin (porcine skin, Sigma) in the absence or presence of 1 μM Fabs for 24 hours at room temperature, then samples were analyzed by 12% SDS-PAGE. A synthetic inhibitor GM6001 and a non-inhibitory Fab R2C17 were used as the positive and negative controls.

Lopez T., Nam D.H., Kaihara E., Mustafa Z, & Ge X. (2017). Identification of Highly Selective MMP-14 Inhibitory Fabs by Deep Sequencing. Biotechnology and bioengineering, 114(6), 1140-1150.

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Evaluation of Rhodamine-Loaded TA-DMNs for Skin Insertion

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To evaluate the skin insertion properties of TA-DMNs, rhodamine B (Rho-B; Sigma-Aldrich, St. Louis, MO, USA) was added to the TA-loaded solution at 0.1% (w/v) to construct needle tips with fluorescence. porcine skins (Cronex, Hwaseong, Korea) stored in a fridge at −20℃ were thawed in PBS for 30 min at room temperature. Thawed porcine skins were wiped and dried to remove the moisture on the surface. Then, Rho-B-loaded TA-DMN patches were applied thoroughly to porcine skin using thumb force or the force exerted by a shooting device and left for 5 min. After that, the patches were peeled off and the needle tips left in the porcine skin were imaged using bright-field microscopy. To assess whether penetration through the skin barrier by TA-DMNs was successful, porcine skin was stained with trypan blue (Sigma-Aldrich, St. Louis, MO, USA) and imaged via bright-field microscopy.

Sim J., Kang G., Yang H., Jang M., Kim Y., Ahn H., Kim M, & Jung H. (2022). Development of Clinical Weekly-Dose Teriparatide Acetate Encapsulated Dissolving Microneedle Patch for Efficient Treatment of Osteoporosis. Polymers, 14(19), 4027.

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Synthesis of Gelatin Methacrylate Hydrogel

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Briefly, a 10% (w/v) gelatine type A solution (porcine skin, 300 bloom strength, Sigma-Aldrich, Gillingham, UK) was dissolved in phosphate-buffered saline (PBS, Sigma-Aldrich, Gillingham, UK) at 60 °C for 30 min. Methacrylic anhydride (MA, Sigma-Aldrich, Gillingham, UK) was added to the gelatine solution to a final concentration of 6% (v/v) and left to react for 2 h at 50 °C under vigorous stirring. After the reaction period, the solution was transferred to 50 mL tubes and the unreacted MA was partially removed by centrifugation at 3500 rpm for 5 min at room temperature. The solution was then dialysed against distilled water for 7 days at 37 °C using 12–14 kDa cut-off dialysis tubes (Thermo Scientific, Paisley, UK). After dialysis, the GelMA solution was diluted to 2% (w/v) and the pH adjusted to 7.4 using 1 mM sodium hydroxide solution (Sigma-Aldrich, Gillingham, UK). Lastly, GelMA solutions were lyophilised for 2 days to generate a white porous foam, which was stored at −80 °C until further use.

Barroso I.A., Man K., Robinson T.E., Cox S.C, & Ghag A.K. (2022). Photocurable GelMA Adhesives for Corneal Perforations. Bioengineering, 9(2), 53.

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