Bovine skin gelatin

Manufactured by Merck Group

Sourced in United States

Bovine skin gelatin is a protein-based material derived from the skin of cattle. It is a versatile laboratory product commonly used in various applications, including as a gelling agent, stabilizer, and emulsifier. The core function of bovine skin gelatin is to provide a consistent and reliable matrix for various experiments and analyses, leveraging its unique physical and chemical properties.

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Lab products found in correlation

Methacrylic anhydride, by Merck Group (3 mentions) Cocl2 6h2o, by Thermo Fisher Scientific (2 mentions) Cuso4 5h2o, by Thermo Fisher Scientific (2 mentions) Glutaric dialdehyde gda, by Thermo Fisher Scientific (2 mentions) Gel pro analyzer software, by Media Cybernetics (1 mentions) Triton x 100, by Merck Group (1 mentions) Alexa fluor 647, by Thermo Fisher Scientific (1 mentions) 2 7 dichlorofluorescin diacetate dcfda, by Merck Group (1 mentions) 3 4 5 dimethylthiazol 2 yl 5 3 carboxymethoxyphenyl 2 4 sulfophenyl 2h tetrazolium mts reagent, by Promega (1 mentions) 3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt reagent, by Merck Group (1 mentions) Zncl2, by Thermo Fisher Scientific (1 mentions) Monochlorobimane mcb, by Thermo Fisher Scientific (1 mentions) Hematoxylin and eosin, by BioVitrum (1 mentions) Cresyl violet acetate, by Thermo Fisher Scientific (1 mentions) Phalloidin cruzfluor 647 conjugate, by Santa Cruz Biotechnology (1 mentions) Recombinant human tgf β1, by R&D Systems (1 mentions) Calcein am, by Thermo Fisher Scientific (1 mentions) Rpmi 1640, by Merck Group (1 mentions) Streptomycin, by Thermo Fisher Scientific (1 mentions) Polycarbonate filter, by Neuro Probe (1 mentions)

Close spelling variants of this product

Gelatin (1674 citations) Bovine gelatin (32 citations) Gelatin from bovine skin (24 citations) Gelatin (type B from bovine skin (17 citations) Type B bovine skin gelatin (10 citations) Bovine skin (6 citations) Bovine skin-derived gelatin (2 citations)

12 protocols using bovine skin gelatin

Gelatin Zymography for MMP-9 and MMP-2 Analysis

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The activity of MMP‐9 and MMP‐2 in the liver tissue from WT and MMP‐13 KO mice was analysed using gelatin zymography 34. About 100 mg of fresh liver tissue was homogenized in 1 ml of 50 mM Tris‐HCl buffer (pH 8.0) containing 150 mM NaCl and 1% Triton X‐100. The homogenates were centrifuged at 5000 × g for 30 min. at 4°C, and the supernatants were collected. After equalizing the protein concentration, 30 μg protein was loaded to a 10% polyacrylamide gel containing 0.1% bovine skin gelatin (#G8150; Sigma‐Aldrich) under non‐reducing conditions. The gels were washed twice (30 min./wash) in 2.5% Triton X‐100 at room temperature and then incubated overnight at 37°C in 50 mM Tris‐HCl buffer (pH 7.6) containing 10 mM CaCl₂. The gels were stained with Coomassie brilliant blue R‐250 for 30 min. and destained until clear transparent bands were developed and photographed. The bands were quantified using Gel‐Pro analyzer software (Media Cybernetics).

George J., Tsutsumi M, & Tsuchishima M. (2017). MMP‐13 deletion decreases profibrogenic molecules and attenuates N‐nitrosodimethylamine‐induced liver injury and fibrosis in mice. Journal of Cellular and Molecular Medicine, 21(12), 3821-3835.

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Synthesis and Purification of GelMA Hydrogel

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Gelatin methacrylate (GelMA) was synthesized as described previously.^{39 (link),40 (link)} Briefly, 10 g of bovine skin gelatin (Sigma Aldrich, St. Louis, MO, USA) was dissolved in 100 mL of PBS, and heated to 60 °C while stirring for an hour until the gelatin fully dissolved. The temperature was lowered to 50 °C, after which 8 mL of methacrylic anhydride (MA; Sigma Aldrich, Cat. No. 276685) was added dropwise to the gelatin solution, which was then stirred vigorously for 1 hour. The solution was diluted with 200 mL of warm PBS, dialyzed against MilliQ ultrapure water using 12–14 kDa cutoff dialysis tubing (Spectrum Laboratories, Rancho Dominguez, CA, USA) for seven days at 40 °C, changing water three times per day, to remove unreacted methacrylic anhydride and contaminants from the solution. The dialyzed GelMA solution was then flash frozen in liquid nitrogen and lyophilized in a freeze dryer for 4 days, and stored at −20 °C.

Agrawal G., Aung A, & Varghese S. (2017). Skeletal muscle-on-a-chip: an in vitro model to evaluate tissue formation and injury. Lab on a chip, 17(20), 3447-3461.

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Cytotoxicity and Oxidative Stress Assays in Skin Cells

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Bovine skin gelatin, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT reagent), 4′,6-diamidino-2-phenylindole (DAPI), phenazine methosulfate (PMS), and Triton X-100 and 2′,7′-dichlorofluorescin diacetate (DCFDA) were purchased from Sigma-Aldrich. Monochlorobimane (MCB) was purchased from ThermoFisher Scientific. CuSO₄·5H₂O, ZnCl₂, CoCl₂·6H₂O, glutaric dialdehyde (GDA), and cresyl violet acetate were obtained from Acros Organics. Citrus pectin (classic CM 201) was obtained from Herbstreith&Fox.
3-(4,5-Dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS reagent) was purchased from Promega. Phalloidin CruzFluor™ 647 conjugate, anti-VEGF (C-1) mouse monoclonal, anti-ICAM-2 (S-16) goat polyclonal, anti-MMP-3 goat monoclonal and anti MMP-3 goat monoclonal antibodies were purchased from Santa Cruz Biotechnology. Anti-HIF-1a mouse monoclonal antibody, donkey anti-mouse IgG (H + L) highly cross-adsorbed secondary antibody, Alexa Fluor 647, and donkey anti-goat IgG (H + L) cross-adsorbed secondary antibody, Alexa Fluor 555, were obtained from ThermoFisher Scientific. Anti-CD31 (PECAM-1) rabbit monoclonal antibody was obtained from Abcam. Hematoxylin and Eosin, and Giemsa staining were purchased from BioVitrum (Russia). Cell culture media and reagents were purchased from Paneco (Russia).

Yergeshov A.A., Zoughaib M., Ishkaeva R.A., Savina I.N, & Abdullin T.I. (2022). Regenerative Activities of ROS-Modulating Trace Metals in Subcutaneously Implanted Biodegradable Cryogel. Gels, 8(2), 118.

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Quantifying Collagen Production in Fibroblasts

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Collagen production was measured using the QuickZyme BioSciences soluble collagen assay kit (Leiden, Netherlands). Fibroblasts (50,000) were grown for 4 d in six-well plates and washed with phosphate-buffered saline (PBS), and collagen was solubilized by adding 0.5 M acetic acid overnight. Measurements were normalized for total protein content. As positive control, cells were exposed for 4 d to 5 ng/ml recombinant human TGFβ1 (R&D Systems, Minneapolis, MN). For zymography, conditioned medium was loaded on a 10% polyacrylamide gel containing 1% bovine skin gelatin (Sigma Aldrich, St. Louis, MO). Gels were incubated for 48 h and then stained with 1% bromophenol blue; subsequently gels were destained with 1% acetic acid.

Marsboom G., Chen Z., Yuan Y., Zhang Y., Tiruppathi C., Loyd J.E., Austin E.D., Machado R.F., Minshall R.D., Rehman J, & Malik A.B. (2017). Aberrant caveolin-1–mediated Smad signaling and proliferation identified by analysis of adenine 474 deletion mutation (c.474delA) in patient fibroblasts: a new perspective on the mechanism of pulmonary hypertension. Molecular Biology of the Cell, 28(9), 1177-1185.

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Investigating GPR55 Signaling in Immune Cells

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Monoclonal IgE (clone SPE7), RPMI 1640, 2-mercaptoethanol (2-ME), Laemmli buffer, L-α-lysophosphatidylinositol (LPI), and sphingosine-1-phosphate (S1P) were purchased from Sigma-Aldrich (St. Louis, MO, USA). HEPES, non-essential amino acids (NEAA), penicillin, streptomycin, and fetal bovine serum (FBS) were obtained from Gibco-BRL-Life Technologies (Gaithersburg, MD, USA). Interleukin (IL)-3 was purchased from PeproTech (Rocky Hill, NJ, USA). The GPR55 synthetic agonist, O-1602, was purchased from Cayman Chemical (Ann Arbor, MI, USA), while the GPR55 antagonist, ML-193, was purchased from TOCRIS (Bristol, UK). The CB2 antagonist, AM630, was from Sigma-Aldrich (St. Louis, MO, USA). Anti-p-cofilin (pSer 3) antibodies were obtained from Cell Signaling Technology (Danvers, MA, USA). Anti-p-LIMK (pThr 508/505) and anti-β-actin (C-terminal) antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). The secondary antibodies, HRP-coupled anti-mouse and anti-rabbit, were obtained from Jackson ImmunoResearch (West Grove, PA, USA). Rhodamine-labeled phalloidin and calcein-AM were obtained from Life Technologies (Carlsbad, CA, USA). DAPI was purchased from Invitrogen (Carlsbad, CA, USA). The polycarbonate filters were from Neuro Probe, and bovine skin gelatin from Sigma-Aldrich (St. Louis, MO, USA)

Martínez-Aguilar L.M., Ibarra-Sánchez A., Guerrero-Morán D.J., Macías-Silva M., Muñoz-Bello J.O., Padilla A., Lizano M, & González-Espinosa C. (2023). Lysophosphatidylinositol Promotes Chemotaxis and Cytokine Synthesis in Mast Cells with Differential Participation of GPR55 and CB2 Receptors. International Journal of Molecular Sciences, 24(7), 6316.

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Perfusion-based Vascular Contrast Imaging

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Mice were deeply anesthetized with 3-fold higher dose of anesthetic (see above). To induce vasodilation, animals were first perfused through the right heart ventricle with the 37^° C warm phosphate buffered saline (PBS with Ca²⁺ and Mg²⁺, Lonza) containing papaverine hydrochloride (4μg/ml, Sigma), adenosine (1mg/ml, Roth) and heparin (50IU/ml, Ratiopharm), followed by infusion of fixative 4% parafolmaldehyde (PFA, Sigma) in PBS. Contrast solution, containing 50% of Bi₃OCl and 5% bovine skin gelatin (both from Sigma) was prepared ahead and pre-warmed to 42⁰C. Mice were first perfused with PBS to remove excess fixative followed by slow infusion of 700μl of the contrast solution, during 2 min. with 120mm Hg pressure. Mice were chilled immediately and immersed in 2% of PFA overnight. For the CT scan (TriFoil imaging eXplore CT 120), the body of the animal was placed into an imaging chamber and positioned with the hind limb region in the center of the field of view. A 360° scan consisting of 1200 single views with a 1 second delay between two views (100 ms, 80 kV, 32 mA) resulted in a voxel size of 50x50x50 µm³. Data were visualized using Osirix viewer software.

Kapanadze T., Bankstahl J.P., Wittneben A., Koestner W., Ballmaier M., Gamrekelashvili J., Krishnasamy K., Limbourg A., Ross T.L., Meyer G.J., Haller H., Bengel F.M, & Limbourg F.P. (2019). Multimodal and Multiscale Analysis Reveals Distinct Vascular, Metabolic and Inflammatory Components of the Tissue Response to Limb Ischemia. Theranostics, 9(1), 152-166.

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Gelatin Methacrylation for Biomaterials

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Gelatin was methacrylated in accordance with the protocol described elsewhere^{31 (link), 32 (link)}. In brief, 10g of bovine skin gelatin (Sigma Aldrich, St. Louis, MO, USA) was dissolved in 100 mL of PBS and stirred at 60 °C for roughly 1 hour to achieve complete solvation. Next the solution was lowered to 50°C, after which, 8mL of methacrylic anhydride (cat no.: 276685; Sigma Aldrich) was added to the solution drop wise with vigorous stirring. The solution was kept at 50°C with vigorous stirring for an hour after the addition was complete, after which, it is quenched with 2x the volume of PBS (200 mL). The solution was then dialyzed against milliQ water using 12–14 kDa cutoff dialysis tubing (Spectrum Laboratories, Rancho Dominguez, CA, USA) for one week (3 times per day water change) at 40 °C to remove trace contaminants. Next, the GelMA solution was frozen in liquid nitrogen and lyophilized in a freeze dryer for 4 days before being stored at −20 °C until usage.

Aung A., Theprungsirikul J., Lim H.L, & Varghese S. (2016). Chemotaxis-driven assembly of endothelial barrier in a tumor-on-a-chip platform. Lab on a chip, 16(10), 1886-1898.

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Synthesis of Gelatin Methacrylate Polymer

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Gelatin methacrylate (GelMA) was synthesized as described earlier.^{38 (link)} Briefly, 10 g of bovine skin gelatin (Sigma Aldrich, St. Louis, MO, USA) was mixed at 10% (wt/v) with 100 ml PBS and stirred at 60°C until fully dissolved. Next, methacrylic anhydride (MA; Sigma Aldrich) was added to the solution at a rate of 0.5 ml/min for a total of 8 ml. The solution was then stirred for 60 minutes at 50°C. After being diluted 2x with warm PBS, the solution was dialyzed against distilled water using 12–14 kDa cutoff dialysis tubing (Spectrum Laboratories, Rancho Dominguez, CA, USA) for one week (3 times/day water change) at 40°C to remove the unreacted methacrylic anhydride and methacrylic acid from the solution. Next, the GelMA solution was frozen using liquid nitrogen and lyophilized in a freeze dryer for 4 days before being stored at −80°C until usage. The dried GelMA was further purified using column chromatography with a Sephadex G-25 column (GE Healthcare Life Sciences, Pittsburgh, PA, USA) and lyophilized.

Aung A., Bhullar I.S., Theprungsirikul J., Davey S.K., Lim H.L., Chiu Y.J., Ma X., Dewan S., Lo Y.H., McCulloch A, & Varghese S. (2015). 3D cardiac μ tissues within a microfluidic device with real-time contractile stress readout. Lab on a chip, 16(1), 153-162.

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Taste Perception and Gustatory Stimuli

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Participants were instructed to rinse with distilled water at least four times over a 1 min period prior to testing. Gelatin was the vehicle for gustatory stimuli. The participants were asked to hold 20 ml gelatin (2% bovine skin gelatin, Sigma-Aldrich, St. Louis, USA) in their mouth with either bitter (0.1% quinine, Sigma-Aldrich, adult toxic dose, 2.5 – 4 g) or neutral taste (2% gelatin) [29 (link)]. The gustatory stimuli were presented at room temperature (~21°C) with an at least four times distilled water rinse after each test for 5 min until the taste has been washed out [30 (link)].

Yang G., Baad-Hansen L., Wang K., Xie Q.F, & Svensson P. (2014). Effect of negative emotions evoked by light, noise and taste on trigeminal thermal sensitivity. The Journal of Headache and Pain, 15(1), 71.

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Ultrastructural Analysis of B Cell Subsets

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For ultrastructural analysis, sorted FSC^loER^lo IgM⁺ B cells from non-stimulated cultures and FSC^hiER^hi and FSC^loER^lo IgM⁺ B cells from LPS-stimulated cultures were sedimented at 300 × g for 5 min and fixed with 4% paraformaldehyde (PFA) (Merck) and 2% glutaraldehyde (GLA) (Electron Microscopy Sciences) in 0.1 M phosphate buffer (PB, pH 7.4) (Merk) for 2 h at RT. Cell pellets were then embedded in 10% bovine skin gelatin (Sigma) and post-fixed with 1% OsO₄ (Sigma), 0.8% K₃ Fe(CN)₆ (Sigma) in water at 4°C for 1 h. Samples were dehydrated with ethanol (Merck) and embedded in epoxy resin (TAAB 812 resin; TAAB Laboratories) according to standard procedures. After polymerization, 80-nm-thick (ultrathin) sections were obtained and stained with uranyl acetate (Electron Microscopy Sciences) and lead citrate (Merck) according to standard procedures. Samples were examined in a JEOL JEM1400Flash electron microscope operating at 100 kV. Images were recorded with an OneView 16-Megapixel CMOS Camera from Gatan. Quantification of cytoplasmic areas in the different B cell subsets was performed using Image J software. At least 30 different cell profiles per condition were analyzed at 1,200X magnification.

Morel E., Herranz-Jusdado J.G., Simón R., Abós B., Perdiguero P., Martín-Martín A., Andrés G., Muñoz-Atienza E., Guerra Rodriguez M., Díaz-Rosales P, & Tafalla C. (2022). Endoplasmic reticulum expansion throughout the differentiation of teleost B cells to plasmablasts. iScience, 26(1), 105854.

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