Leghon Red Eggs Commercial Egg

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Lipoprotein isolation and characterization: Leghon red hens commercial eggs ( > 73g.) were used for this study. The shell was broken, the white of an egg was eliminated and the yolk was put on a filter paper to roll in it to eliminate completely the remainders of the white adhered. The yolk membrane was punctured and the content of the yolk (approximately 15 ml) was deposited in a test tube. An equivalent volume of saline solution was added (NaCl 0.16 M; pH = 7) and it was mixed completely. The resultant solution was centrifugated for 30 minutes at 9000 rpm at 4ºC in a rotor Beckman JA-20, eliminating the pellet. The supernatant was collected to obtain the lipoproteins by gradient density ultracentrifugation: 10ml of the yolk solution were taken and then 0.9 g of potassium bromide (KBr) was added. The bromide was dissolved with a smooth agitation, with care not to denature the proteins. Straight afterwards, saline solution was added (NaCl 0.16 M, pH=7) to the 10ml of the yolk solution with a Pasteur pipette, avoiding the sample diffusion, forming two phases and filling the tube completely. The ultracentrifugation was carried out during 19.5 h at 4ºC and 45000 rpm. (244.500 x g) in a Kontron Centrikon T-2190 ultracentrifuge in a TFT 50.38 rotor,…show more content…
Their visualisation was performed using Coomassie brilliant blue (G-250). GS-800TM Calibrated Densitometer (BioRad, California, USA) was used to scan the gels and the resulting image file was assayed using the Quantity One analysing software (BioRad, California, USA). Molecular masses were estimated according to Precision Plus Protein unstained standards (BioRad, California, USA) which contains ten highly purified recombinant proteins with molecular masses from 10 to 250

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