Insulin from porcine pancreas

Manufactured by Merck Group

Sourced in Switzerland, United States

Insulin from porcine pancreas is a laboratory product derived from the pancreas of pigs. It is a protein hormone that plays a key role in regulating blood sugar levels. The product is primarily used for research and analytical purposes in various scientific and medical applications.

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Lab products found in correlation

Cryschem plate, by Hampton Research (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Chicken egg white lysozyme, by Merck Group (1 mentions)

Close spelling variants of this product

Insulin (4936 citations) Porcine insulin (43 citations) Porcine pancreas (18 citations)

3 protocols using insulin from porcine pancreas

Insulin Crystallization Protocol

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Insulin from porcine pancreas (catalogue No. I5523) was purchased from Sigma–Aldrich GmbH, Buchs, Switzerland and dissolved in 50 mM Na₂HPO₄, 10 mM EDTA pH 10.5 to a final concentration of 25 mg ml⁻¹. Crystals were grown using two different methods for different parts of the investigation. The crystals used in Sections 3.1 and 3.4 were grown in batch by mixing 0.5 ml of the insulin solution in a 1:1 ratio with the crystallization buffer [25%(w/v) PEG 6000, 0.1 M bis-Tris propane pH 7.5, 0.2 M sodium bromide] in a centrifuge tube at 20°C. The centrifuge tube was briefly vortexed and left on a revolver/rotator at 20°C. The mean crystal size and concentration were 30 µm and 1.5 × 10⁶, respectively. The crystals used in Sections 3.2 and 3.3 were grown overnight at 20°C in 24-well sitting-drop vapor-diffusion CrysChem plates (Hampton Research, USA) against 500 µl reservoir. The drops contained 2 µl insulin solution and 2 µl crystallization buffer.

Martiel I., Beale J.H., Karpik A., Huang C.Y., Vera L., Olieric N., Wranik M., Tsai C.J., Mühle J., Aurelius O., John J., Högbom M., Wang M., Marsh M, & Padeste C. (2021). Versatile microporous polymer-based supports for serial macromolecular crystallography. Acta Crystallographica. Section D, Structural Biology, 77(Pt 9), 1153-1167.

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Cell Culture Media and Reagents

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Media [Dulbecco’s modified Eagles medium (DMEM) low Glucose (5.5 mM) with Glutamax], PBS (including Ca²⁺ and Mg²⁺), antibiotics and heat inactivated sera (fetal bovine serum–FBS and horse serum—HS) were purchased from Gibco Life Technologies (Grand Island, NY, USA). Insulin from porcine pancreas (Sigma Aldrich Chemical Co., St. Louis, MO, USA) was dissolved in dilute acetic acid according to manufacturer recommendations and kept frozen at -20°C. Metabolic inhibitors of: PI3-K (LY294002), MAPK MEK (PD98059), GSK-3β (SB216763 and LiCl), ETC complex I (rotenone), mitochondrial ATP synthase complex (oligomycin) if necessary were dissolved in DMSO and depending on the type were kept frozen at -20°C or refrigerated at 4–8°C. All other reagents were cell culture tested, of high purity and unless otherwise stated they were purchased from Sigma-Aldrich Chemical Co. (St. Louis, MO, USA). Plastics were from Becton Dickinson (BD Biosciences, Franklin Lakes, NJ, USA), tubes for deep freezing from NunclonTM (Nunc^TM, Roskilde, Denmark) while syringe filters were purchased from Corning-Costar Inc. (Cambridge, MA, USA).

Litwiniuk A., Pijet B., Pijet-Kucicka M., Gajewska M., Pająk B, & Orzechowski A. (2016). FOXO1 and GSK-3β Are Main Targets of Insulin-Mediated Myogenesis in C2C12 Muscle Cells. PLoS ONE, 11(1), e0146726.

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On-Chip Protein Crystallization via Dialysis

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Chicken egg-white lysozyme was purchased from Sigma Aldrich as a lyophilized powder, dissolved in distilled water and filtered to obtain a solution with a final concentration of ∼30 mg mL -1 . Insulin from porcine pancreas was also purchased from Sigma Aldrich as a lyophilized powder, dissolved in 0.5 M hydrochloric acid at pH 3 and filtered to obtain a solution with a final concentration of ∼2.5 mg mL -1 . IspE, a kinase from Agrobacterium tumefaciens, was used with a final concentration of ∼10 mg mL -1 (F. Borel et al. 48 ). All protein solutions were filtered through 0.22 μm Millipore filters prior to their use. The membranes integrated into the chip were the standard regenerated cellulose (RC) membranes Spectra/ Por® (Spectrum Labs). The crystallization mixtures were all centrifuged and filtered to remove all solid particles (precipitate, dust or nuclei of crystals) prior to their use. Table 1 summarizes the experimental conditions (protein concentration, initial concentration of the crystallization solution, MWCO of the membrane, and temperature) for the on-chip crystallization of lysozyme, IspE and insulin through the dialysis method.

Junius N., Jaho S., Sallaz-Damaz Y., Borel F., Salmon J.B, & Budayova-Spano M. (2020). A microfluidic device for both on-chip dialysis protein crystallization and in situ X-ray diffraction. Lab on a chip, 20(2).

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