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    Wyświetlanie 1-7 z 7
    Tytuł:
    Application of electrophoretic methods for detection of protein-porphyrin complexes.
    Autorzy:
    Kowalska, Agnieszka
    Kwiek, Piotr
    Miłosz, Ewa
    Gondek, Grzegorz
    Romiszewska, Anna
    Graczyk, Alfreda
    Podhajska, Anna
    Powiązania:
    https://bibliotekanauki.pl/articles/1043407.pdf
    Data publikacji:
    2003
    Wydawca:
    Polskie Towarzystwo Biochemiczne
    Tematy:
    porphyrins
    photodynamic therapy
    PDT
    electrophoresis
    Opis:
    Simple methods for detection and isolation of protein-porphyrin complexes were elaborated in our laboratory. They are based on the separation of protein-porphyrin complexes in native polyacrylamide gel and measurement of their fluorescence, with the use of two detection systems: the commercially available Gel DocTM 2000 system, and a system specially designed for the purpose of these investigations, concerning protein-porphyrin interactions. The fluorescent complexes can be electro-transferred from the gel onto PVDF membrane, eluted and analyzed in order to identify the protein interacting with porphyrins.
    Źródło:
    Acta Biochimica Polonica; 2003, 50, 4; 1155-1163
    0001-527X
    Pojawia się w:
    Acta Biochimica Polonica
    Dostawca treści:
    Biblioteka Nauki
    Artykuł
    Tytuł:
    Native nucleic acid electrophoresis as an efficient alternative for genotyping method of influenza virus
    Autorzy:
    Pajak, Beata
    Lepek, Krzysztof
    Powiązania:
    https://bibliotekanauki.pl/articles/1039250.pdf
    Data publikacji:
    2014
    Wydawca:
    Polskie Towarzystwo Biochemiczne
    Tematy:
    gel electrophoresis
    SSCP
    influenza
    minor genetic variants
    Opis:
    Influenza viruses are the worldwide major causative agents of human and animal acute respiratory infections. Some of the influenza subtypes have caused epidemics and pandemics among humans. The varieties of methods are available for the rapid isolation and identification of influenza viruses in clinical and environmental samples. Since nucleic acids amplification techniques such as RT-PCR have been adapted, fast and sensitive influenza type and subtype determination is possible. However, in some ambiguous cases other, more detailed assay might be desired. The genetic material of influenza virus is highly unstable and constantly mutates. It is known that single nucleotide polymorphisms (SNPs) results in resistance to commercially available anti-viral drugs. The genetic drift of the virus could also result in weakening of immune response to infection. Finally, in a substantial number of patients co-infection with various virus strains or types has been confirmed. Although the detection of co-infection or presence of minor genetic variants within flu-infected patients is not a routine procedure, a rapid and wide spectrum diagnostics of influenza virus infections could reveal an accurate picture of the disease and more importantly, is crucial for choosing the appropriate therapeutics and virus monitoring. Herein we present the evidences that native gel electrophoresis and MSSCP - a method based on multitemperature single strand conformation polymorphism could furnish a useful technique for minor variants, which escape discovery by conventional diagnostic assays.
    Źródło:
    Acta Biochimica Polonica; 2014, 61, 3; 479-483
    0001-527X
    Pojawia się w:
    Acta Biochimica Polonica
    Dostawca treści:
    Biblioteka Nauki
    Artykuł
    Tytuł:
    Using capillary electrophoresis to study methylation effect on RNA-peptide interaction.
    Autorzy:
    Mucha, Piotr
    Szyk, Agnieszka
    Rekowski, Piotr
    Agris, Paul
    Powiązania:
    https://bibliotekanauki.pl/articles/1043465.pdf
    Data publikacji:
    2003
    Wydawca:
    Polskie Towarzystwo Biochemiczne
    Tematy:
    methylated nucleosides
    methylation
    arginine methylation
    RN-peptide interaction
    capillary electrophoresis
    Opis:
    Methylation of RNA and proteins is one of a broad spectrum of post-transcriptional/translational mechanisms of gene expression regulation. Its functional signification is only beginning to be understood. A sensitive capillary electrophoresis mobility shift assay (CEMSA) for qualitative study of the methylation effect on biomolecules interaction is presented. Two RNA-peptide systems were chosen for the study. The first one consists of a 17-nucleotide analogue (+27-+43) of the yeast tRNAPhe anticodon stem and loop domain (ASLPhe) containing three of the five naturally occurring modifications (2'-O-methylcytidine (Cm32), 2'-O-methylguanine (Gm34) and 5-methylcytidine (m5C40)) (ASLPhe-Cm32,Gm34,m5C40) and a 15-amino-acid peptide (named tF2 : Ser1-Ile-Ser-Pro-Trp5-Gly-Phe-Ser-Gly-Leu10-Leu- Arg-Trp-Ser-Tyr15) selected from a random phage display library (RPL). A peptide-concentration-dependent formation of an RNA-peptide complex was clearly observable by CEMSA. In the presence of the peptide the capillary electrophoresis (CE) peak for triply methylated ASLPhe shifted from 18.16 to 20.90 min. Formation of the complex was not observed when an unmethylated version of ASLPhe was used. The second system studied consisted of the (+18)-(+44) fragment of the trans-activation response element of human immunodeficiency virus type 1 (TAR RNA HIV-1) and a 9-amino-acid peptide of the trans-activator of transcription protein (Tat HIV-1) Tat(49-57)-NH2 (named Tat1 : Arg49-Lys-Lys-Arg52-Arg-Gln-Arg-Arg- Arg57-NH2). In the presence of Tat(49-57)-NH2 a significant shift of migration time of TAR from 18.66 min to 20.12 min was observed. Methylation of a residue Arg52→Arg(Me)2, crucial for TAR binding, strongly disrupted formation of the complex. Only at a high micromolar peptide concentration a poorly shaped, broad peak of the complex was observed. CE was found to be an efficient and sensitive method for the analysis of methylation effects on interaction of biomolecules.
    Źródło:
    Acta Biochimica Polonica; 2003, 50, 3; 857-864
    0001-527X
    Pojawia się w:
    Acta Biochimica Polonica
    Dostawca treści:
    Biblioteka Nauki
    Artykuł
    Tytuł:
    A proteomics approach to identify the differential protein level in cardiac muscle of diabetic rat
    Autorzy:
    Karthik, Dhanaraj
    Vijayakumar, Ravichandran
    Pazhanichamy, Kalailingam
    Ravikumar, Sivanesan
    Powiązania:
    https://bibliotekanauki.pl/articles/1039290.pdf
    Data publikacji:
    2014
    Wydawca:
    Polskie Towarzystwo Biochemiczne
    Tematy:
    diabetes mellitus
    cardiac muscle proteome
    2D electrophoresis
    MALDI-TOF-MS
    phylogenetic analysis
    Opis:
    Background: Cardiovascular proteomics investigation reveals the characterization and elucidation of the novel therapeutic targets and strategies to prevent the development of heart failure associated diabetic complication by using 2DE and MS. Methods: The experimental animals were made diabetic with a single intraperitoneal injection of alloxan (150 mg/kg of bw). Albino rats were randomly divided into four individual groups: Group-I control (n=6), group-II alloxan-induced diabetic rats, untreated (n=6), group-III (n=6) and group-IV (n=6) alloxan-induced diabetic rats were treated with aqueous and ethanolic extracts of Cynodon dactylon for 15 days, respectively. Animals were euthanized to collect the heart tissues and blood samples. 2DE sample preparation, gel running and staining (n=6: each groups) were performed at the same time to avoid variation. The result of six gel images from each group were analyzed and evaluated as one match set with 2D software (P<0.05). Results: The above experiment revealed two up-regulated proteins in group-II i.e. NTF4 and ETFB. Conclusions: NTF4 is a neuro-protective agent for neuro-degenerative diseases. It will prevent diabetic secondary complications, such as diabetic polyneuropathy and cardiomyopathy. ETFB is active in the mitochondria, the energy-producing centres in cells. It is clear from the experiment that because of up-regulation of ETFB more energy is availabile and the electron transfer for heart during diabetes is possible, what leads to reduce the oxidative stress and free-radical formation. The up-regulated proteins reduced CVD that occurred just before overt hyperglycaemia due to administration of C. dactylon. This approach established the preliminary reference map for decoding cellular mechanisms linked between pathogenesis CVD and diabetes.
    Źródło:
    Acta Biochimica Polonica; 2014, 61, 2; 285-293
    0001-527X
    Pojawia się w:
    Acta Biochimica Polonica
    Dostawca treści:
    Biblioteka Nauki
    Artykuł
    Tytuł:
    Molecular basis of cellulose biosynthesis disappearance in submerged culture of Acetobacter xylinum
    Autorzy:
    Krystynowicz, Alina
    Koziołkiewicz, Maria
    Wiktorowska-Jezierska, Agnieszka
    Bielecki, Stanisław
    Klemenska, Emilia
    Masny, Aleksander
    Płucienniczak, Andrzej
    Powiązania:
    https://bibliotekanauki.pl/articles/1041378.pdf
    Data publikacji:
    2005
    Wydawca:
    Polskie Towarzystwo Biochemiczne
    Tematy:
    2-D electrophoresis
    UDP-glucose pyrophosphorylase
    phosphoglucomutase
    bacterial cellulose
    Acetobacter xylinum
    PCR-MP
    Opis:
    Acetobacter xylinum strains are known as very efficient producers of bacterial cellulose which, due to its unique properties, has great application potential. One of the most important problems faced during cellulose synthesis by these bacteria is generation of cellulose non-producing cells, which can appear under submerged culture conditions. The reasons of this remain unknow. These studies have been undertaken to compare at the molecular level wild-type, cellulose producing (Cel+) A. xylinum strains with Cel- forms of cellulose-negative phenotype. Comparison of protein profiles of both forms of A. xylinum by 2D electrophoresis allowed for the isolation of proteins which were produced exclusively by either Cel+ or Cel- cells. Sequences of peptides derived from these proteins were aligned with those of proteins deposited in databases. This analysis revealed that Cel- cells lacked two enzymes: phosphoglucomutase and glucose-1-phosphate uridylyltransferase, which generates UDP-glucose being the substrate for cellulose synthase. DNA was analyzed by ligation-mediated PCR carried out at low denaturation temperature (PCR-MP). Two DNA fragments of different thermal stability (218 and 217 bp) were obtained from the DNA of Cel+ and Cel- forms, respectively. The only difference between these Cel- and Cel+ DNA fragments is deletion of one T residue. Alignment of those two sequences with those deposited in the GenBank database revealed that similar fragments are present in the genomes of some bacterial cellulose producers and are located downstream from open reading frames (ORF) encoding phosphoglucomutase. The meaning of this observation is discussed.
    Źródło:
    Acta Biochimica Polonica; 2005, 52, 3; 691-698
    0001-527X
    Pojawia się w:
    Acta Biochimica Polonica
    Dostawca treści:
    Biblioteka Nauki
    Artykuł
    Tytuł:
    Comparative proteomic analysis of Bombyx mori hemolymph and fat body after calorie restriction
    Autorzy:
    Chen, Huiqing
    Li, Yijia
    Chen, Keping
    Yao, Qin
    Li, Guohui
    Wang, Lin
    Powiązania:
    https://bibliotekanauki.pl/articles/1040312.pdf
    Data publikacji:
    2010
    Wydawca:
    Polskie Towarzystwo Biochemiczne
    Tematy:
    calorie restriction
    Bombyx mori
    two-dimensional gel electrophoresis
    proteomic analysis
    MALDI-TOF/TOF MS
    Opis:
    Calorie restriction (CR) is known to extend life span from yeast to mammals. To gain an insight into the effects of CR on growth and development of the silkworm Bombyx mori at protein level, we employed comparative proteomic approach to investigate proteomic differences of hemolymph and fat body of the silkworm larvae subjected to CR. Thirty-nine differentially expressed proteins were identified by MALDI TOF/TOF MS. Among them, 19 were from the hemolymph and 20 from the fat body. The hemolymph of the CR group contained two down-regulated and 17 up-regulated proteins, whereas the fat body contained 15 down-regulated and five up-regulated ones. These proteins belonged to those functioning in immune system, in signal transduction and apoptosis, in regulation of growth and development, and in energy metabolism. Our results suggest that CR can alter the expression of proteins related to the above four aspects, implying that these proteins may regulate life span of the silkworm through CR.
    Źródło:
    Acta Biochimica Polonica; 2010, 57, 4; 505-511
    0001-527X
    Pojawia się w:
    Acta Biochimica Polonica
    Dostawca treści:
    Biblioteka Nauki
    Artykuł
    Tytuł:
    Distinct expression, localization and function of two Rab7 proteins encoded by paralogous genes in a free-living model eukaryote
    Autorzy:
    Osińska, Magdalena
    Wiejak, Jolanta
    Wypych, Emilia
    Bilski, Henryk
    Bartosiewicz, Rafał
    Wyroba, Elżbieta
    Powiązania:
    https://bibliotekanauki.pl/articles/1039860.pdf
    Data publikacji:
    2011
    Wydawca:
    Polskie Towarzystwo Biochemiczne
    Tematy:
    glycosylation
    antipeptide antibodies
    Real-Time PCR
    Rab7 isotypes
    RNAi
    2D electrophoresis
    Paramecium octaurelia
    STED
    Opis:
    Rab7 GTPases are involved in membrane trafficking in the late endosomal/lysosomal pathway. In Paramecium octaurelia Rab7a and Rab7b are encoded by paralogous genes. Antipeptide antibodies generated against divergent C-termini recognize Rab7a of 22.5 kDa and Rab7b of 25 kDa, respectively. In 2D gel electrophoresis two immunoreactive spots were identified for Rab7b at pI about 6.34 and about 6.18 and only one spot for Rab7a of pI about 6.34 suggesting post-translational modification of Rab7b. Mass spectrometry revealed eight identical phosphorylated residues in the both proteins. ProQ Emerald staining and ConA overlay of immunoprecipitated Rab7b indicated its putative glycosylation that was further supported by a faster electrophoretic mobility of this protein upon deglycosylation. Such a post-translational modification and substitution of Ala140 in Rab7a for Ser140 in Rab7b may result in distinct targeting to the oral apparatus where Rab7b associates with the microtubular structures as revealed by STED confocal and electron microscopy. Rab7a was mapped to phagosomal compartment. Absolute qReal-Time PCR analysis revealed that expression of Rab7a was 2.6-fold higher than that of Rab7b. Upon latex internalization it was further 2-fold increased for Rab7a and only slightly for Rab7b. Post-transcriptional gene silencing of rab7a suppressed phagosome formation by 70 % and impaired their acidification. Ultrastructural analysis with double immunogold labeling revealed that this effect was due to the lack of V-ATPase recruitment to phagolysosomes. No significant phenotype changes were noticed in cells upon rab7b silencing. In conclusion, Rab7b acquired a new function, whereas Rab7a can be assigned to the phagolysosomal pathway.
    Źródło:
    Acta Biochimica Polonica; 2011, 58, 4; 597-607
    0001-527X
    Pojawia się w:
    Acta Biochimica Polonica
    Dostawca treści:
    Biblioteka Nauki
    Artykuł
      Wyświetlanie 1-7 z 7

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