SDS–PAGE

SDS - PAGE
  • 文章类型: Journal Article
    重组基于弹性蛋白的蛋白质(ELPs)用于包括治疗的应用,药物输送,和组织工程由于其生物相容性和独特的能力进行简单的凝聚。这里,我们描述了一种经济有效的方法来纯化ELPs利用盐沉淀和它们的可逆相变性质时,加热到其较低的临界溶解温度(LCST)。此外,我们描述了用于粘合剂应用的将酪氨酸残基转化为L-3,4-二羟基苯丙氨酸(DOPA)的翻译后修饰。
    Recombinant elastin-based proteins (ELPs) are used in applications that include therapeutics, drug delivery, and tissue engineering due to their biocompatibility and unique ability to undergo simple coacervation. Here, we describe a cost-effective method to purify ELPs utilizing salt precipitation and their reversible phase transition property when heated above their lower critical solution temperature (LCST). Furthermore, we describe the post-translational modification of converting tyrosine residues to L-3,4-dihydroxyphenylalanine (DOPA) for adhesive applications.
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  • 文章类型: Journal Article
    用于太阳能收集和转换的可生物降解和可持续无毒材料的设计和生产是一项重大挑战。这里,我们的目标是报告新型蛋白质/脂质水凝胶的制备,并证明它们在两项正交基础研究中的实用性-光捕获和白光发射。我们的水凝胶含有高达90%的水,同时也可以用注射器独立注射。在一个应用中,我们用合适的有机供体-受体染料加载这些水凝胶,并证明了四种不同染料之间的能量转移级联,以最红色的染料作为能量目的地。我们假设染料作为单体实体嵌入远离水池的蛋白质/脂质相中,并且四种染料中的任何一种的激发都会导致最低能量受体的强烈发射。与能量转移级联相反,我们展示了使用这些凝胶来形成发白光的水凝胶染料组件,其中激励迁移受到严重约束。通过限制染料到染料的能量转移,蓝色,绿色,和红色染料在各自的波长发射,从而产生复合白光发射。发射的CIE颜色坐标为0.336和0.339-几乎是纯的白光发射。因此,两个具有相反要求的相关研究可以容纳在同一个水凝胶中,它是通过一种简单的方法由可食用的成分制成的。这些凝胶在释放到环境中时是可生物降解的,可持续,并且可能对能量应用感兴趣。
    The design and production of biodegradable and sustainable non-toxic materials for solar-energy harvesting and conversion is a significant challenge. Here, our goal was to report the preparation of novel protein/lipid hydrogels and demonstrate their utility in two orthogonal fundamental studies-light harvesting and white-light emission. Our hydrogels contained up to 90% water, while also being self-standing and injectable with a syringe. In one application, we loaded these hydrogels with suitable organic donor-acceptor dyes and demonstrated the energy-transfer cascade among four different dyes, with the most red-emitting dye as the energy destination. We hypothesized that the dyes were embedded in the protein/lipid phase away from the water pools as monomeric entities and that the excitation of any of the four dyes resulted in intense emission from the lowest-energy acceptor. In contrast to the energy-transfer cascade, we demonstrate the use of these gels to form a white-light-emitting hydrogel dye assembly, in which excitation migration is severely constrained. By restricting the dye-to-dye energy transfer, the blue, green, and red dyes emit at their respective wavelengths, thereby producing the composite white-light emission. The CIE color coordinates of the emission were 0.336 and 0.339-nearly pure white-light emission. Thus, two related studies with opposite requirements could be accommodated in the same hydrogel, which was made from edible ingredients by a simple method. These gels are biodegradable when released into the environment, sustainable, and may be of interest for energy applications.
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  • 文章类型: Journal Article
    从Boswelliaserrata(Bs)和Salixalba(Sa)中提取的提取物用作家禽饲料中的补充剂。这项研究的目的是研究膳食补充Bs和Sa提取物对血清和蛋白的可能影响。锌和铁,和蛋黄胆固醇含量在开始产蛋的关键阶段来角母鸡。总共120个小母鸡,17周龄,分为两组(对照组(C)和治疗组(T),每个n=60)。T组接受含有Bs(5%)和Sa(5%)的补充剂12周。这项研究持续了19周。使用琼脂糖凝胶电泳(AGE)和SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)分级分离血清蛋白。采用原子吸收光谱法测定血清中的微量元素,用比色试验测定蛋黄胆固醇。对于分析的生化指标,在对照和补充母鸡之间没有观察到显著差异。此外,补充植物提取物不会对血清蛋白的生理变化产生负面影响;因此,它可以安全地用作治疗,以防止在发病和早期铺设阶段的炎症状态。
    Extracts from Boswellia serrata (Bs) and Salix alba (Sa) are used as supplements in poultry feed. The aims of this research were to study the possible effects of dietary supplementation with Bs and Sa extracts on serum and albumen proteins, zinc and iron, and yolk cholesterol content in Leghorn hens during the critical phase of the onset of laying. A total of 120 pullets, 17 weeks of age, were assigned to two groups (control (C) and treated (T), n = 60 each). The T group received a supplement containing Bs (5%) and Sa (5%) for 12 weeks. The study lasted 19 weeks. Serum proteins were fractionated using agarose gel electrophoresis (AGE) and SDS-polyacrylamide gel electrophoresis (SDS-PAGE). Trace elements were determined in serum using atomic absorption spectrometry, and yolk cholesterol was determined using a colorimetric test. No significant differences were observed between control and supplemented hens for the analyzed biochemical indices. Moreover, the supplementation with phytoextracts did not negatively affect the physiological variations in serum proteins; therefore, it can be safely used as a treatment to prevent inflammatory states at onset and during the early laying phase.
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  • 文章类型: Journal Article
    用九种不同的蛋白酶制剂进行羽扇豆蛋白分离物(LPI;羽扇豆)的酶水解,以研究其对技术功能性的影响,感官特性,和蛋白质的完整性来估计免疫反应性的降低。Alcalase2.4L,木瓜蛋白酶,和胃蛋白酶在通过SDS-PAGE分析检查的α-和β-conglutin的降解中最有效,虽然水解度仅略有增加。LPI溶解度的技术功能特性,乳化,大多数蛋白水解酶改善了泡沫活性,从LPI的980%泡沫活性提高到胃蛋白酶水解产物的3,614%泡沫活性。苦味的形成,最有可能与苦味肽的产生有关,在碱性蛋白酶水解物中很明显,而其他水解产物与LPI相比没有显示出苦味的大量增加。除Alcalase处理外,水解产物的其他感官属性也与LPI非常相似。这项研究的结果表明,LPI的酶促降解具有修饰IgE反应多肽并改善分离物的技术功能的潜力,因此可以将其用作食品成分。
    Enzymatic hydrolysis of lupin protein isolates (LPI; Lupinus angustifolius L.) was performed with nine different protease preparations to investigate their effect on technofunctionality, sensory properties, and the integrity of the proteins to estimate the reduction of the immunoreactivity. Alcalase 2.4 L, papain, and pepsin were most effective in the degradation of the α- and β-conglutin examined by SDS-PAGE analysis, although the degree of hydrolysis only slightly increased. The technofunctional properties of LPI-solubility, emulsifying, and foaming activity-were improved by most of the proteolytic enzymes with the most impressive increase from 980% foam activity for LPI up to 3,614% foam activity for pepsin hydrolysate. The formation of bitterness, most likely linked to generation of bitter peptides, was pronounced in the Alcalase hydrolysate, while the other hydrolysates did not show an extensive increase in bitterness compared to the LPI. Other sensory attributes of the hydrolysates-with the exception of Alcalase treatment-were also very similar to the LPI. The results of this study show the potential of enzymatic degradation of LPI to modify the IgE-reacting polypeptides and to improve the technofunctionality of the isolates and therefore their use as food ingredients.
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  • 文章类型: Journal Article
    The present work was conducted to evaluate the quality of milk (fresh/acidic/neutralized) on the physico-chemical, textural and fatty acid profile of khoa prepared from buffalo milk and stored in poly-alu-poly laminates for 30 °C/7 days and 5 °C/21 days, respectively. The degree of deterioration of common quality parameters was rapid during storage at 30 °C as compared to storage at 5 °C. Khoa stored at 30 °C showed greater variation in various physico-chemical and textural parameters as compared to khoa stored at 5 °C. Acidity, ash, tyrosine value, furosine, HMF, FFA, peroxide value, TBA value, butyric acid and stearic acid showed an increasing trend whereas, decrease in pH and oleic acid was observed as storage period progressed. Noticeable changes were observed in textural attributes of khoa during storage. However, the SDS-PAGE pattern of caseins from different types of khoa showed almost negligible deviation during storage.
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  • 文章类型: Journal Article
    Gamma radiation is a very effective tool for inducing genetic variation in characters of many plants. Black seeds of M2 mutant were obtained after exposure of an Egyptian cowpea cultivar (Kaha 1) to a low dose of gamma rays. Segregation of seed coat color, weight of 100 seeds and seed eye pattern of the black seeds of this mutant line were further examined in this study. Four colors were observed for seed coat in the M3 plants ranging from cream to reddish brown and three eye patterns were distinguished from each other. SDS-PAGE of the seed storage proteins showed 18 protein bands; five of these bands disappeared in the seeds of M3 plants compared to M2 and M0 controls while other 5 protein bands were specifically observed in seeds of M3 plants. PCR analysis using twelve ISSR primers showed 47 polymorphic and 8 unique amplicons. The eight unique amplicons were characteristic of the cream coat color and brown wide eye pattern (M03-G10) while the polymorphic bands were shared by 6 coat-color groups. A PCR fragment of 850 bp was amplified, using primer HB-12, in M3-G04 which showed high-100 seed weight. These results demonstrated the mutagenic effects of gamma rays on seed coat color, weight of 100 seeds and eye pattern of cowpea M3 mutant plants.
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  • 文章类型: Journal Article
    Somatic chromosome study from root tip cells using the squash technique of the cytological method and seed protein profile of 5 varieties of Lens culinaris (Lentil) through SDS-PAGE were investigated. Karyotype analysis showed gross uniformity in morphology. Somatic chromosome number 2n = 14 is constant for all the varieties. Chromosomes are mostly long to medium in length with secondary constrictions in one pair of chromosome. Primary constrictions in chromosome ranged from nearly median to nearly submedian in most of the cases. Notwithstanding the gross homogeneity, karyotype analysis revealed minute differences in details. Each lentil variety is thus characterized by its own karyotype, serving as one of the identifying criteria. The seed protein profile revealed that varieties are very close to each other with respect to similarity index that ranged from 0.594 to 0.690. With regard to seed protein banding patterns, slight polymorphism (14.285%) indicating low genetic diversity has been identified among the 5 varieties. A dendrogram indicates one variety is plesiomorphic and rest varieties are apomorphic. All the experimental varieties of lentil studied here show low genetic diversity due to their similar genetic base, indigenous genetic resources and conservative nature of the seed protein.
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  • 文章类型: Journal Article
    Pathological α-synuclein (α-syn) overexpression and iron (Fe)-induced oxidative stress (OS) are involved in the death of dopaminergic neurons in Parkinson\'s disease (PD). We have previously characterized the role of triacylglycerol (TAG) formation in the neuronal response to Fe-induced OS. In this work we characterize the role of the α-syn variant A53T during Fe-induced injury and investigate whether lipid metabolism has implications for neuronal fate. To this end, we used the N27 dopaminergic neuronal cell line either untransfected (UT) or stably transfected with pcDNA3 vector (as a transfection control) or pcDNA-A53T-α-syn (A53T α-syn). The overexpression of A53T α-syn triggered an increase in TAG content mainly due to the activation of Acyl-CoA synthetase. Since fatty acid (FA) β-oxidation and phospholipid content did not change in A53T α-syn cells, the unique consequence of the increase in FA-CoA derivatives was their acylation in TAG moieties. Control cells exposed to Fe-induced injury displayed increased OS markers and TAG content. Intriguingly, Fe exposure in A53T α-syn cells promoted a decrease in OS markers accompanied by α-syn aggregation and elevated TAG content. We report here new evidence of a differential role played by A53T α-syn in neuronal lipid metabolism as related to the neuronal response to OS.
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  • 文章类型: Journal Article
    Zymography, the detection of proteolytic activities on the basis of protein substrate degradation, has been a technique described in the literature for at least in the past 50 years. In this study, we used substrate immersing zymography to analyze proteolysis of proteases. Instead of being directly added into a sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gel, the substrates were added into the immersing solution after electrophoresis. With substrate immersing zymography, some characters of proteases, such as enzyme forms, potential proteolytic activity, molecular weights, presence of complexes, and potentially active enzyme fragments in complex biological samples, can be determined.
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  • 文章类型: Journal Article
    Antioxidant properties of ethanol extract of Silybum marianum (milk thistle) seeds was investigated. We have also investigated the protein damage activated by oxidative Fenton reaction and its prevention by Silybum marianum seed extract. Antioxidant potential of Silybum marianum seed ethanol extract was measured using different in vitro methods, such as lipid peroxidation, 1,1-diphenyl-2-picrylhydrazyl (DPPH) and ferric reducing power assays. The extract significantly decreased DNA damage caused by hydroxyl radicals. Protein damage induced by hydroxyl radicals was also efficiently inhibited, which was confirmed by the presence of protein damage markers, such as protein carbonyl formation and by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The present study shows that milk thistle seeds have good DPPH free radical scavenging activity and can prevent lipid peroxidation. Therefore, Silybum marianum can be used as potentially rich source of antioxidants and food preservatives. The results suggest that the seeds may have potential beneficial health effects providing opportunities to develop value-added products.
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