SDS对肉仔鸡肌肉和哺乳动物肝组织蛋白质提取的效果

Syeda Qandiel Zahra, Sidra Latif, H. Nazir, Zunaira Izhar Shah, Azka Zafar, A. Majid, Adil Farooq, A. Qureshi
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摘要

背景:本研究旨在验证十二烷基硫酸钠(SDS)作为一种合适且经济高效的裂解缓冲液的潜力,以最大限度地从肉鸡和哺乳动物肝脏的各种肌肉组织中提取蛋白质。材料与方法:三种不同的肌肉组织(I;胸部,二世;翼和III;随机选取商品肉鸡(n=4),提取腿部组织,解剖小鼠(n=3),提取肝脏组织样本。SDS的比例为1:1 (w/v);肌肉组织和肝脏组织的最佳提取时间分别为10%和1.0 & 1.5%。孵育后,各组织匀浆,离心。用溴甲酚绿(BCG)法提取粗蛋白(CP)。结果:SDS(10%)孵育1小时后CP产率最高。检测sds试剂对肌肉组织类型和孵育时间的共依赖性,发现组织I(胸部)在孵育1小时后CP含量最高,组织II(翅膀)在孵育3小时后CP含量较多,组织III(腿部)在孵育1、2和3小时后CP含量相等。从哺乳动物肝组织中,以1.5%的SDS得到CP (6.9 g/dl)和ALB (1.6 g/dl)的最大产量。1.0% SDS均质后未检测CP和白蛋白(Alb)含量。差异有统计学意义(P< 0.05)。结论:在肉鸡肌肉组织中,10%的SDS是提取粗蛋白质的最佳缓冲液浓度,而在小鼠肝脏样品中,1.5%的SDS比1.0%的SDS更好。此外,这种简单和最便宜的程序和易于制备的试剂可能适用于提取重要的组织蛋白组分。
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Efficacy of SDS For Protein Extraction from Broiler Muscles and Mammalian Liver Tissue
Background: The present study purports to check and validate the potential of sodium dodecyl sulfate (SDS) alone being a suitable and cost-effective lysis buffer for maximum and efficient protein extraction from various muscle tissues of broiler chicken and mammalian liver. Materials and Methods: Three different muscle tissues (I; chest, II; wing and III; leg) were extracted from randomly selected commercial broilers (n=4) while mice (n=3) were dissected for the extraction of liver tissue samples. 1:1 ratio (w/v) of SDS; 10 and 1.0 & 1.5% was used for muscles and liver tissues, respectively for its best time optimization for protein extraction. After incubation, respective tissues were homogenized followed by centrifugation. The supernatant was then processed for crude protein (CP) extraction by Bromocresol Green (BCG) method. Results: SDS (10%) achieved a maximum yield of CP after 1 hour of incubation. When checked the co-dependence of SDS-reagent on muscle-tissue type and time of incubation, tissue I (chest) was found to give maximum CP contents after 1 hour of incubation, tissue II (wing) extracted more CP after 3 hours while tissue III (leg) rendered equal amounts of CP after 1, 2 and 3 hours of incubation, respectively. From the mammalian liver tissue maximum yield of CP (6.9 g/dl), and albumin (ALB) (1.6 g/dl) was obtained with 1.5% of SDS. While the CP and albumin (Alb) content was not detected after homogenization with 1.0% SDS. Significance was checked at (P< 0.05). Conclusion: It is concluded from the above findings that 10% SDS is the best lysis buffer concentration to extract crude protein from all the studied broiler muscle tissues while from mice liver samples we found 1.5% SDS lysis reagent seems good than 1.0%. Furthermore, this simple and cheapest procedure and ease of preparation this reagent may be suitable for extraction of important tissue protein fractions.
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