scholarly journals Karakterisasi Protein Lernaea Cyprinacea Dengan Metode Elektroforesis SDS-PAGE [Characterization Of Protein Lernaea cyprinacea By Using SDS-PAGE Electrophoresis Method]

2019 ◽  
Vol 2 (1) ◽  
pp. 61
Author(s):  
Gunanti Mahasri, Ulia Fajriah, Sri Subekti

Abstract One of the diseases that often attack are fish is parasitic disease caused by Lernaea cyprinacea, called Lernaeosis. This ectoparasites could be detected on skin, gill, eyes, fin or inside of mouth and nostril of fishes. The aim of the study was to know the character is tic of L.cyprinacea protein based on molecular weight using SDS-PAGE. The results of this study were expected as Lernaeosis vaccine candidate. The experiment used SDS-PAGE with 12% separating gel, 3% stacking gel, voltage 110 volts 28 A and gel staining using coomassie blue. The result showed that, there were eight protein bands with molecular weight 82.3, 73.3, 66.6, 60.5, 54.9, 27.5, 23.1 and 19.8 kDa. Among them, the bands with molecular weight 27.5, 23.1 and 19.8 kDa were thicker and clearer than the others.

1986 ◽  
Vol 34 (2) ◽  
pp. 209-214 ◽  
Author(s):  
J U Alles ◽  
K Bosslet

A new monoclonal antibody (moab BW 200) of IgG3 kappa-isotype was generated which recognizes an epitope located on an antigen molecule restricted to human neoplastic and non-neoplastic endothelial cells. The molecular weight of the antigen was determined using immunoprecipitation analysis followed by SDS-PAGE. Despite its similar molecular weight to FVIII-RAG, the antigen detected by moab BW 200 was shown to be different from FVIII-RAG.


1981 ◽  
Author(s):  
Ellinor I Peerschke ◽  
Mariorie B Zucker ◽  
Avner Rotman

The interaction of fibrinogen with its, platelet membrane receptor was investigated using 125-labeled fibrinogen which was photoaffinity labeled with a light-sensitive azide. This photoreactive material (125I-NPA-fibr) was indistinguishable from unlabeled fibrinogen as well as from iodinated fibrinogen on SDS-PAGE. It bound specifically to platelets stimulated with ADP and was crosslinked to the platelet membrane after exposure to light ( λ >300 nm) for 4 min. No crosslinking was observed in the presence of EDTA or with platelets that failed to aggregate with ADP either due to the congenital deficiency thrombasthenia or following incubation with EDTA for 8 min at 37° , pH 7.8 and recalcification. SDS-PAGE of platelets bearing crosslinked 125I-NPA-fibr revealed a radiolabeled band of about 450,000 daltons in addition to the 340,000 dalton radioactive band of fibrinogen, suggesting that fibrinogen had been covalently bound to a platelet membrane component with an intact apparent molecular weight of approximately 110,000 daltons. Following reduction, an extra radioactive band was noted at 80,000 daltons. As the A∝-chains of fibrinogen were too weakly labeled to be detected by autoradiography, this indicated that either the Bβ or γchain of fibrinogen was attached to a 25,000-35,000 molecular weight platelet membrane fragment. We conclude that the additional radioactive bands observed after electrophoresis of platelets bearing specifically bound-photoaffinity labeled 125I-fibrinogen most likely represent the binding of the B β or γ chains of fibrinogen to the platelet fibrinogen receptor which may be GPIIb.


Author(s):  
D.W. Estry ◽  
T.G. Bell ◽  
G.H. Tishkoff ◽  
J.C. Mattson ◽  
S.C. Estry

A protein analogous to human antithrombin III was isolated from fresh horse plasma. The procedure for purification was a modification of Thaler and Schmer’s two-step isolation procedure. The horse protein was homogeneous on 7.5% SDS-PAGE gels and had a molecular weight of 62,000 to 64,000 daltons in both reducing and non-reducing systems (human; 62,300). Rabbit anti-human antithrombin III was used to demonstrate a line of partial identity by Immunoelectrophoresis between the horse and human protein. The horse protein rapidly neutralizes human thrombin (34,000 daltons) and the reaction appears to be greatly potentiated by heparin. In order to establish the formation of 1:1 covalent stoichiometric complex between horse AT III and thrombin (IIa), time studies were run in the presence and absence of heparin. AT III (62,000) at 15 seconds, 2, 5, 10 and 60 minutes formed a stable complex with thrombin (32,000) having a molecular weight of 86,000 daltons. Additional bands developing with time are due to the autolytic capabilities of the uncomplexed IIa. The major autolytic band had a molecular weight of 70,000 daltons. Addition of heparin potentiated the interaction although it did not change the stoichio-metry of the complexes formed. The data accumulated to date demonstrates the similarities between the human and horse protein and the possibilities of using the horse as a model system for the evaluation of AT III replacement therapy in vivo.


1996 ◽  
Vol 38 (6) ◽  
pp. 401-406 ◽  
Author(s):  
Yano Tomomasa ◽  
Cleide Ferreira Catani ◽  
Michiko Arita ◽  
Takeshi Honda ◽  
Toshio Miwatani

The mannose-resistant hemagglutinating factor (HAF) was extracted and purified from a diffuse adherent Escherichia coli (DAEC) strain belonging to the classic enteropathogenic E. coli (EPEC) serotype (0128). The molecular weight of HAF was estimated to be 18 KDa by SDS-PAGE and 66 KDa by Sephadex G100, suggesting that the native form of HAF consists of 3-4 monomeric HAF. Gold immunolabeling with specific HAF antiserum revealed that the HAF is not a rigid structure like fimbriae on the bacterial surface. The immunofluorescence test using purified HAF on HeLa cells, in addition to the fact that the HAF is distributed among serotypes of EPEC, suggests that HAF is a possible adhesive factor of DAEC strains


2015 ◽  
Vol 18 (2) ◽  
pp. 151
Author(s):  
Sebastian Margino ◽  
J. Jumi'ati ◽  
N. Ngadiman

Potato Cyst Nematode (PCN), Globodera rostochiensis, is one of the important potato’s pests and causedeconomic looses up to 70% in the several centrals of potato plantations in Indonesia. PCN’s shell componentof egg shell containing chitin (inner layer) and viteline/ protein (outer layer). The purpose of this researchwas to purify of protease Bacillus sp. TBRSN-1, isolate from tomato’s rhizosfer in Yogyakarta province. Thepurifi ed protease could be used for cutting the life cycle of PCN. Results showed that Bacillus sp. TBRSN-1could produce extracellular protease and purifi cation using DEAE-cellulose ion-exchange chromatographyand Sephacryl S-300 gel fi ltration chromatography resulted in specifi c activity 4.31 fold and 1.68% recovery.Analysing using SDS-PAGE 12.5% and molecular weight 48.1 kDa. Km and Vmax values of the protease forcasein substrate were 7.83 mg/ml and 4.03 μg/h, respectively. The optimum activity at the temperature30oC and pH 7.0. Keywords : protease, purifi cation, indigenous Bacillus sp. TBRSN-1


2019 ◽  
Vol 2 (1) ◽  
pp. 126
Author(s):  
Suardi Suardi ◽  
Ana Hidayanti Mukarromah ◽  
Stalis Norma Ethica

Fish is a potential source of animal protein, but has a weakness that is easy to rot. To avoid decay can be preserved by salting the fish. In this study wet salting was carried out to analyze the effect of salting on fish protein. The sample used was a type of gourami with 5 tails. 1 for the sample before salting and 4 for salting each salted at 10, 20, 30 and 40% b/v after that it was left to stand for 12 hours. The research method used was the Gel Electrophoresis method (SDS-PAGE) to determine molecular weight (MW), looking at the purity and damage of proteins in the sample. The samples of gouramy before salting showed 16 bands, 8 major bands and 8 minor ribbons. Samples of gouramy with a salt concentration of 10% b/v showed 16 bands, 7 major bands and 9 minor bands. Samples of gouramy with a salt concentration of 20% b/v showed 14 bands, 7 major bands and 7 minor bands. Samples of gouramy with a salt concentration of 30% b/v showed 10 bands, 3 major bands and 7 minor bands. Samples of gouramy with a salt concentration of 40% b/v showed 9 bands, 3 major bands and 6 minor bands. Thus it can be concluded that salting of fish can affect the gouramy protein, which is the higher the salt content added, the protein found in the fish will be denatured. The salting process of 10% b/v in gouramy meat is the most recommended salting process compared to the salting process of 20, 30 and 40% b/v.Ikan merupakan sumber protein hewani yang potensial, namun memiliki suatu kelemahan yaitu mudah membusuk. Untuk menghindari pembusukan dapat dilakukan pengawetan dengan penggaraman pada ikan. Pada penelitian ini dilakukan penggaraman basah untuk menganalisis pengaruh penggaraman terhadap protein ikan. Sampel yang digunakan adalah jenis ikan gurami sebanyak 5 ekor. 1 ekor untuk sampel sebelum penggaraman dan 4 ekor dilakukan penggaraman masing-masing digarami dengan kadar 10, 20, 30 dan 40% b/v setelah itu didiamkan selama 12 jam. Metode penelitian yang digunakan adalah metode Elektroforesis Gel (SDS- PAGE) untuk menentukan berat molekul (BM), melihat kemurnian dan kerusakan protein pada sampel. Pada sampel ikan gurami sebelum penggaraman menunjukkan 16 pita, 8 pita mayor dan 8 pita minor. Sampel ikan gurami dengan konsentrasi garam 10% b/v menujukkan 16 pita, 7 pita mayor dan 9 pita minor. Sampel ikan gurami dengan konsentrasi garam 20% b/v menunjukkan 14 pita, 7 pita mayor dan 7 pita minor. Sampel ikan gurami dengan konsentrasi garam 30% b/v menunjukkan 10 pita, 3 pita mayor dan 7 pita minor. Sampel ikan gurami dengan konsentrasi garam 40% b/v menunjukkan 9 pita, 3 pita mayor dan 6 pita minor. Dengan demikian dapat disimpulkan bahwa penggaraman pada ikan dapat berpengaruh terhadap protein ikan gurami yaitu makin tinggi kadar garam yang ditambahkan maka protein yang terdapat pada ikan akan terdenaturasi. Proses penggaraman 10% b/v pada daging ikan gurami merupakan proses penggaraman yang paling disarankan dibandingkan proses penggaraman 20,30 dan 40% b/v.


Author(s):  
Amiruddin Amiruddin ◽  
Tongku Nizwan Siregar ◽  
Amalia Sutriana ◽  
Dwinna Aliza ◽  
T. Armansyah

This study has long-term objectives to obtain immunogenic prototype that can be used to induce multiple ovulation in goats. Working steps of this study were begun with the collection of ovarium from goats, collection of granulose cells, culture of granulose and characterization of molecular weight and isoelectric point (pI) of inhibin protein of granulose cells obtained from culture and non-culture of granulose cells, and followed by preparation of monoclonal antibody toward inhibin. The results showed that inhibin isolated either from culture or non-culture of granulose cells produced a 32 kDa band. Molecular weight of inhibin was measured by Western Blot. The 32 kDa band of SDS PAGE product appeared on Western Blot result was inhibin molecules produced by granulose cells collected fom culture and non-culture of granulose cells that can be identified by Mab-inhibin. Product of IEF gel electrophoresis suggested that inhibin molecule collected from culture of granulose cells has no charge at isoelectric points ranging from 5-6, depends on its total amino acid composition.


ALCHEMY ◽  
2019 ◽  
Vol 7 (1) ◽  
pp. 7 ◽  
Author(s):  
Dewi Yuliani ◽  
Dhienda Risa Awalsasi ◽  
Akyunul Jannah

<p>Gelatin, a proteinaceous additive, is obtained from hydrolysis of collagen in the bone, hide and skin of animals. As natural product, gelatin has been applied in many industries with various functions. This study attempt to characterize gelatin profile of broiler chicken (<em>Gallus domestica</em>) using SDS-PAGE electrophoresis. The chicken bone was pretreated using a strong base, sodium hydroxide, producing type B gelatin. The gelatin was purified through precipitation using the variation of ammonium sulfate concentrations (40-70%) and dialysis using cellophane membrane. The purified gelatin was characterized through SDS-PAGE electrophoresis. Based on electrophoresis visualization, reduction of band intensity by ammonium sulfate 40% showed removal of small peptide fragments. The remained gelatin showed two major bands, α-chains and a β-chain with the respective molecular weight of ~135 and ~245 kDa. The protein content of the unpurified gelatin (E1) was 71.65±0.60 mg/L.  The purified E1 gelatins by 40-70% of ammonium sulfate addition contained 61.42±3.90, 60.45±1.36, 59.89±0.24, and 55.32±1.05 mg/L of protein concentration, respectively.</p><p> </p><p>Keywords: chicken bone, gelatin profile, protein electrophoresis</p>


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