Detection of UVR8 Homodimers and Monomers by Immunoblotting Analysis in

2021 ◽  
pp. 83-93
Author(s):  
Guoqian Yang ◽  
Xiaorui Liu ◽  
Li Lin
Keyword(s):  
Immunoblotting Analysis

Immune responses of Texel sheep to excretory/secretory products of adult Haemonchus contortus

Parasitology ◽  
1994 ◽  
Vol 108 (3) ◽  
pp. 351-357 ◽  
Author(s):  
H. D. F. H. Schallig ◽  
M. A. W. van Leeuwen ◽  
W. M. L. Hendrikx
Keyword(s):  
Immune Response ◽  
Immune Responses ◽  
Haemonchus Contortus ◽  
Lymphocyte Proliferation ◽  
The Other ◽  
Proliferation Index ◽  
Serum Samples ◽  
Immunoblotting Analysis ◽  
Molecular Weights ◽  
Secretory Products

SUMMARYThe excretory/secretory (E/S) products of adult Haemonchus contortus comprise of at least 15 polypeptides with molecular weights ranging from 10 to > 100 kDa. These E/S products induce an immune response in infected Texel sheep, as demonstrated by specific IgGI levels and a significant lymphocyte proliferation index. Moreover, immunoblotting analysis revealed that sera of primary H. contortus-infected sheep specifically recognize a 24 kDa E/S product. In addition, sera of challenged sheep react strongly with a 15 kDa E/S product. The other E/S products of H. contortus showed immunoreactivity with serum samples of Haemonchus-infected sheep as well as with samples of sheep harbouring other trichostrongylid infections. These cross-reacting epitopes are the main cause of the lack of specificity of an E/S material- based ELISA. This ELISA can differentiate Haemonchus infections from Nematodirus battus infections, but not from Ostertagia circumcincta or Trichostrongylus colubriformis infections.

Immunoblotting analysis of wheat allergens: control of side reactions through wheat polypeptides naturally present in dried cow milk

2010 ◽  
Vol 21 (3) ◽  
pp. 237-251 ◽  
Author(s):  
Isabelle Bouchez-Mahiout ◽  
Elodie Boulenc ◽  
Jacques Snégaroff ◽  
Dominique Choudat ◽  
Catherine Pecquet ◽  
...  
Keyword(s):  
Cow Milk ◽  
Side Reactions ◽  
Immunoblotting Analysis

Purification and Characterization of Factor VII Inhibitor Found in a Patient with Life Threatening Bleeding

2000 ◽  
Vol 83 (01) ◽  
pp. 60-64 ◽  
Author(s):  
Seiji Miyamoto ◽  
Atsushi Iwasa ◽  
Masao Ishii ◽  
Kenji Okajima ◽  
Yu-ichi Kamikubo
Keyword(s):  
Light Chain ◽  
Calcium Ion ◽  
Polyacrylamide Gel Electrophoresis ◽  
Factor Vii ◽  
Immunoblotting Analysis ◽  
Calcium Dependent ◽  
Sds Page ◽  
Carboxyglutamic Acid ◽  
Life Threatening ◽  
Gla Domain

SummaryWe recently observed a patient with acquired inhibitor-induced F.VII deficiency whose plasma level of F.VII was < 1.0%. However, the biochemical nature of the inhibitor has not yet been clarified. In the present study, we purified the F.VII inhibitor from the patient’s plasma by using activated F.VII (F.VIIa)-conjugated gel and characterized the inhibitor. The results showed that the inhibitor comprised two kinds of antibodies: one was eluted with EDTA (antibody 1) and the other with glycine-HCl buffer (pH 2.3) (antibody 2) from the F.VIIa affinity gel. SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting analysis of these inhibitors demonstrated that both antibodies had features of immunoglobulin G1 (IgG1) with κ and λ-light chains. Antibody 1 bound to the immobilized F.VIIa with a high affinity in the presence of calcium ion, while antibody 2 bound to the F.VIIa very weakly and the binding was independent of calcium ion. Immunoblotting analysis demonstrated that antibody 1 bound to the light chain of F.VIIa after reduction with 2-mercaptoethanol, while it did not react with either the γ carboxyglutamic acid (Gla)-domainless light chain of F.VIIa or the heavy chain with the protease domain. Antibody 1 markedly inhibited the activity of tissue factor-F.VIIa complex. Based on these observations, it is suggested that F.VIIa autoantibody (antibody 1) recognizes the calcium-dependent conformation within or near the Gla domain and inhibits F.VIIa activity by interacting with the light chain.

scholarly journals Molecular characterization and Immunodiagnostic potential of various antigenic proteins of Fasciola gigantica species isolated from sheep of North West Himalayan Region

2019 ◽  
Vol 56 (2) ◽  
pp. 93-107
Author(s):  
J. S. Dar ◽  
B. A. Ganai ◽  
R. A. Shahardar ◽  
U. R. Zargar
Keyword(s):  
Small Ruminants ◽  
Fasciola Gigantica ◽  
Ovis Aries ◽  
Digenetic Trematode ◽  
Diagnostic Specificity ◽  
Immunoblotting Analysis ◽  
Accurate Identification ◽  
North West ◽  
Specificity And Sensitivity ◽  
Sds Page

SummaryThe control of the digenetic trematode Fasciola gigantica has been the major challenge in both cattle and small ruminants as there is a paucity of an effective and commercial vaccine. Thus, the accurate identification and prepatent diagnosis of F. gigantica is an essential prerequisite for its successful prevention and control. In the present study, the morphologically identified specimens isolated from the liver and bile ducts of sheep (Ovis aries) were validated through molecular data. The sequence analysis of ITS-2 of our isolates showed high degree of similarity with F. gigantica and F. hepatica using BLAST function of NCBI. The phylogenetic analysis of our isolates showed a close relationship with previously described F. gigantica and F. hepatica isolates from different countries. The antigenic profile of somatic and E/S antigens of F. gigantica were revealed by SDS–PAGE and immunoblotting using sera from sheep naturally infected with F. gigantica. By SDS-PAGE, 20 distinct bands were revealed from crude somatic fraction. Immunoblotting analysis of these proteins with positive sera exhibited 8 sero-reactive bands ranging from 14 to 97 kDa. Among these 38 and 44 kDa bands were quite specific with high diagnostic specificity and sensitivity. The E/S fraction comprised 7 distinct bands, as revealed by SDS-PAGE analysis. Immunoblotting analysis of these proteins with positive sera exhibited 6 antigenic bands ranging from 23 – 54 kDa. Among these 27 and 33 kDa were found to be quite specific with high diagnostic specificity and sensitivity. The present study concludes that the protein bands of 38 and 44 kDa in somatic fraction and 27 and 33 kDa in E/S fraction can be used for the immunodiagnostic purpose for this economically important parasite, which may also entice further studies regarding their vaccine potential.

scholarly journals Proteomic Characterization of a Mouse Model of Familial Danish Dementia

2012 ◽  
Vol 2012 ◽  
pp. 1-8 ◽  
Author(s):  
Monica Vitale ◽  
Giovanni Renzone ◽  
Shuji Matsuda ◽  
Andrea Scaloni ◽  
Luciano D'Adamio ◽  
...  
Keyword(s):  
Ion Trap ◽  
Scaffold Protein ◽  
Synaptic Function ◽  
Differentially Expressed Proteins ◽  
Wild Type ◽  
Dominant Mutation ◽  
Immunoblotting Analysis ◽  
Familial Danish Dementia ◽  
Dual Specificity

A dominant mutation in theITM2B/BRI2gene causes familial Danish dementia (FDD) in humans. To model FDD in animal systems, a knock-in approach was recently implemented in mice expressing a wild-type and mutant allele, which bears the FDD-associated mutation. Since theseFDDKImice show behavioural alterations and impaired synaptic function, we characterized their synaptosomal proteome via two-dimensional differential in-gel electrophoresis. After identification by nanoliquid chromatography coupled to electrospray-linear ion trap tandem mass spectrometry, the differentially expressed proteins were classified according to their gene ontology descriptions and their predicted functional interactions. The Dlg4/Psd95 scaffold protein and additional signalling proteins, including protein phosphatases, were revealed by STRING analysis as potential players in the altered synaptic function ofFDDKImice. Immunoblotting analysis finally demonstrated the actual downregulation of the synaptosomal scaffold protein Dlg4/Psd95 and of the dual-specificity phosphatase Dusp3 in the synaptosomes ofFDDKImice.

The role of lamin LIII in nuclear assembly and DNA replication, in cell-free extracts of Xenopus eggs

1991 ◽  
Vol 98 (3) ◽  
pp. 271-279
Author(s):  
J. Meier ◽  
K.H. Campbell ◽  
C.C. Ford ◽  
R. Stick ◽  
C.J. Hutchison
Keyword(s):  
Dna Replication ◽  
Membrane Structures ◽  
Chromatin Decondensation ◽  
Immunoblotting Analysis ◽  
Nuclear Assembly ◽  
Contrast Microscopy ◽  
Egg Extracts ◽  
Xenopus Egg ◽  
Xenopus Egg Extracts

Xenopus egg extracts, which support nuclear assembly and DNA replication, were functionally depleted of lamin LIII by inoculating them with monoclonal anti-lamin antibodies. Phase-contrast microscopy and electron-microscopy studies indicated that lamin-depleted extracts supported efficient chromatin decondensation, and assembly of double membrane structures and nuclear pores on demembranated sperm heads. Immunofluorescence microscopy suggests that lamin-antibody complexes are transported across the nuclear membrane but do not assemble into a lamina. These findings were confirmed by immunoblotting analysis of isolated nuclei. Metabolic labelling studies with either biotin-11-dUTP or [32P]dCTP, revealed that nuclei lacking a lamina were unable to initiate DNA replication and that, although such nuclei could import proteins required for DNA replication (e.g. PCNA), these proteins were apparently not organized into replicon clusters.

Anticancer Drug Action on Poly(A) Polymerase Activity and Isoforms during Hela and Wish Cell Apoptosis

2000 ◽  
Vol 15 (2) ◽  
pp. 171-178 ◽  
Author(s):  
N.A.A. Balatsos ◽  
M. Havredaki ◽  
C.M. Tsiapalis
Keyword(s):  
Anticancer Agents ◽  
Enzyme Assay ◽  
Polymerase Activity ◽  
Activity Levels ◽  
Immunoblotting Analysis ◽  
A Cell ◽  
Lower Mobility ◽  
Cell Commitment ◽  
Diploid Cells ◽  
Mrna Polyadenylation

Poly(A) polymerase (PAP; EC 2.7.7.19) catalyzes mRNA polyadenylation. Its activity and isoform levels vary during cell cycle transformation and apoptosis. It has become widely accepted that cell death after DNA damage by anticancer agents is primarily the result of apoptosis and that cells able to evade apoptosis will be resistant to cell killing. The therapeutic agents interferon (IFN), 5-fluorouracil (5-FU) and tamoxifen (Tam) with different mechanisms of action mediate both partial dephosphorylation and inactivation of PAP, detected by immunoblotting analysis and PAP enzyme assay, respectively. We examined the apoptotic tendencies of HeLa and WISH cell lines caused by one of the drugs used, 5-FU. The trend in the cells examined, observed by DAPI and/or DNA fragmentation assay, was found to be accompanied by and reversibly related to PAP activity levels and PAP lower mobility phosphorylated forms of 106 and 100 kDa isoforms. Moreover, a cell type-modulated, differential response of HeLa (chemosensitive cells) versus WISH (drug-resistant diploid cells) has been revealed. This finding yields information on the possible use of PAP as a tumor marker involved in cell commitment and/or induction of apoptosis and may help to improve our understanding of tumor cell sensitivity to anticancer agents.

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