Erythrocyte Immobilisation with Total Protein Extracted from Gnetum gnemon L. Seeds on Variety of Solid Media

Hery Haryanto, S. Nurliana, Syariffuddin Syarifuddin
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Abstract

The objective of this research was to determine the effectiveness of erythrocyte immobilisation with total protein extracted from gnetum seeds on a variety of solid media, i.e., filter papers, doff plastics, and nitrocellulose membranes. Old gnetum seeds were ground to make powders. The powders were homogenized in ml PBS buffer containing anti-protease PMSF in a cool room. The homogenate was centrifuged in order to separate supernatant from pellet. Solid media were punched with a perforator, so the results were discs of papers, plastics, and nitrocellulose membranes. The solid media perforated discs were then dipped in gnetum total protein supernatants. The dipped discs were air-dried for a while. Human erythrocytes of all ABO groups were washed three times and diluted to suspension of 20% (v/v) in NaCl 0,9% (w/v). A small drop of erythrocyte suspension was put in middle of each solid media disc, and its blood suspension was spread evenly. Next, the disc was observed under an USB digital microscope to see whether there was erythrocyte immobilisation on solid media coated with total proteins of gnetum seeds or not. All the erythrocytes from human ABO groups were bound effectively, from strongest to weakest,to paper filters, nitrocellulose membranes, and plastics. The future use of erythrocyte immobilisation on a variety of solid media coated with of gnetum total protein seed may be useful for diagnosing diseases related patological erythrocytes with a specific marker.
木耳种子总蛋白在多种固体培养基上固定化红细胞的研究
本研究的目的是确定从芒草种子中提取的总蛋白在各种固体介质上固定红细胞的有效性,即滤纸、脱胶和硝化纤维素膜。旧的亚麻籽被磨成粉末。粉末在含有抗蛋白酶PMSF的ml PBS缓冲液中均质,在凉爽的房间中。将匀浆离心,从球团中分离上清。固体介质用穿孔器打孔,因此结果是纸、塑料和硝化纤维膜的圆盘。然后将固体介质穿孔盘浸入总蛋白上清液中。浸渍的圆盘被风干了一会儿。所有ABO血型的人红细胞洗涤三次,在NaCl 0,9% (w/v)中稀释至20% (v/v)的悬液。在每个固体介质圆盘中间滴入少量红细胞悬液,使其血悬液均匀分布。接下来,在USB数码显微镜下观察圆盘,观察是否有红细胞固定在包被麻子总蛋白的固体培养基上。所有来自人类ABO血型的红细胞,从最强到最弱,都能有效地与纸过滤器、硝化纤维素膜和塑料结合。将来,在各种涂有总蛋白种子的固体培养基上固定化红细胞,可能有助于用特异性标记物诊断与疾病相关的病理红细胞。
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