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IgG2, Bovine; ELISA

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[#YSRTPBP004] IgG2, Bovine; ELISA


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(1) Systemic humoral immunity in beef bulls following therapeutic vaccination against Tritrichomonas foetus.[TOP]

Pubmed ID :29773139
Publication Date : //
The utility of therapeutic vaccination of bulls against Tritrichomonas foetus has been advocated in previous studies, but anecdotal reports suggest this practice does not clear infections and may additionally confound diagnostic testing by reducing parasite burdens below detectable limits. The objective of this study was to characterize the systemic humoral immune response to therapeutic vaccination in T. foetus-infected bulls over a period of four months using an indirect ELISA and to compare the dynamics of this response to culture and PCR results to establish the existence of a relationship (or lack thereof) between immunization and infection status. A study population of 4- to 6-year-old T. foetus-infected beef bulls (n = 20) was divided equally into a treatment group and a control group. The treatment group received two doses of commercially prepared whole cell killed vaccine 2 weeks apart while the control group received injections of vaccine diluent. Blood samples were collected at each injection and at 4 subsequent dates every 4 weeks thereafter (i.e. 0, 2, 6, 10, 14, and 18 wks) to measure IgG and IgG antibody subisotype response via an indirect ELISA. Preputial smegma samples were collected at the four monthly intervals following vaccination for diagnosis of infection via InPouch™ culture, Modified Diamond's Medium (MDM) culture, and PCR. Humoral response for both IgG isotypes from week 2 through week 18 were significantly increased in vaccinates compared to controls. No significant decrease in infection prevalence was detected in the treatment group for any of the diagnostic methods used. The apparent lack of pathogen clearance during a stimulated immune response suggests that therapeutic vaccination may not be a useful T. foetus management practice.

Authors : Alling Christopher, Rae D Owen, Ma Xiaojie, Neumann Laura, Lollis L Gene, Steele Elizabeth, Yelvington John, Naikare Hemant K, Walden Heather Stockdale, Crews John, Boughton Raoul,

(2) Two-Dimensional MoS-Based Zwitterionic Hydrophilic Interaction Liquid Chromatography Material for the Specific Enrichment of Glycopeptides.[TOP]

Pubmed ID :29742898
Publication Date : //
Mass spectrometry (MS)-based glycoproteomics research requires highly efficient sample preparation to eliminate interference from non-glycopeptides and to improve the efficiency of glycopeptide detection. In this work, a novel MoS/Au-NP (gold nanoparticle)-L-cysteine nanocomposite was prepared for glycopeptide enrichment. The two-dimensional (2D) structured MoS nanosheets served as a matrix that could provide a large surface area for immobilizing hydrophilic groups (such as L-cysteine) with low steric hindrance between the materials and the glycopeptides. As a result, the novel nanomaterial possessed an excellent ability to capture glycopeptides. Compared to commercial zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC) materials, the novel nanomaterials exhibited excellent enrichment performance with ultrahigh selectivity and sensitivity (approximately 10 fmol), high binding capacity (120 mg g), high enrichment recovery (more than 93%), satisfying batch-to-batch reproducibility, and good universality for glycopeptide enrichment. In addition, its outstanding specificity and efficiency for glycopeptide enrichment was confirmed by the detection of glycopeptides from an human serum immunoglobulin G (IgG) tryptic digest in quantities as low as a 1:1250 molar ratio of IgG tryptic digest to bovine serum albumin tryptic digest. The novel nanocomposites were further used for the analysis of complex samples, and 1920 glycopeptide backbones from 775 glycoproteins were identified in three replicate analyses of 50 μg of proteins extracted from HeLa cell exosomes. The resulting highly informative mass spectra indicated that this multifunctional nanomaterial-based enrichment method could be used as a promising tool for the in-depth and comprehensive characterization of glycoproteomes in MS-based glycoproteomics.

Authors : Xia Chaoshuang, Jiao Fenglong, Gao Fangyuan, Wang Heping, Lv Yayao, Shen Yehua, Zhang Yangjun, Qian Xiaohong,

(3) Assembly of Multifunctionalized Gold Nanoparticles with Chemiluminescent, Catalytic, and Immune Activity for Label-Free Immunoassays.[TOP]

Pubmed ID :29727158
Publication Date : //
In this study, we report a universal label-free immunoassay to detect antigen based on multifunctionalized gold nanoparticles (MF-GNPs), which were obtained by successive assembly of N-aminobutyl- N-ethylisoluminol functionalized gold nanoparticles (ABEI-GNPs) with antibody, bovine serum albumin (BSA) and Co. MF-GNPs exhibited excellent chemiluminescent (CL), catalytic and immune activity. It was demonstrated that the CL signal of MF-GNPs decreased in the presence of antigens via antigen-antibody specific binding using human immunoglobulin G (hIgG) and corresponding antibody goat anti-human IgG (anti-hIgG) as a model system, due to that immunoreaction led to the aggregation of GNPs. According to the decreased CL intensity, hIgG could be determined in the range of 1.0 fM to 1.0 nM with a low detection limit of 0.13 fM. Furthermore, this CL strategy was also confirmed to be a general one by replacing hIgG with heart-type fatty acid-binding protein (H-FABP), which is a biomarker of early acute myocardial infarction (AMI). The CL strategy could be employed to detect H-FABP ranging from 10.0 fM to 10.0 nM, and the detection limit is 7.8 fM. The CL strategy also showed good selectivity. It might be extended to detect other antigens if their corresponding antibodies are available.

Authors : Huang Yao, Gao Lingfeng, Cui Hua,

(4) Evaluation of the humoral immune response to a multicomponent recombinant vaccine against S. aureus in healthy pregnant heifers.[TOP]

Pubmed ID :29704938
Publication Date : //
Staphylococcus aureus is a worldwide pathogen that causes mastitis in dairy herds. Shortcomings in control programs have encouraged the development of vaccines against this pathogen. This study evaluated the vaccine candidate VacR, which included recombinant S. aureus protein clumping factor A (rClf), fibronectin binding protein A (rFnBP) and hemolysin beta (rBt), formulated with a novel immune-stimulating complex. Comparisons were made between healthy pregnant heifers that received either VacR (n=8; VacR group) or phosphate buffered saline (PBS) plus adjuvant (control group) SC in the supramammary lymph node area on days 45 and 15 before the expected calving date. Blood and foremilk samples were collected from 7 to 60days post-calving. After calving, heifers in the VacR group produced higher total IgG (IgG) titers against each component, in both serum (rBt, 3.4×10; rClf, 3.1×10; rFnBP, 2.3×10) and milk (rBt, 2.6×10; rClf, 1.3×10; rFnBP, 1.1×10), than control heifers (P<0.0001). There were increased concentrations of IgG and IgG in VacR group (P<0.05), in both serum and milk. Humoral responses remained high throughout the period most susceptible to intramammary infections (P<0.01). Antibodies produced against S. aureus rClf and rFnBP reduced bacterial adherence to fibronectin and fibrinogen by 73% and 67%, respectively (P<0.001). Milk antibodies against these adhesins inhibited S. aureus invasion of a mammary epithelial cell line (MAC-T), resulting in 15.7% of bacteria internalized (P<0.0001). There was an approximately 6-fold reduction in the hemolysis titer for the native hemolysin in the VacR group compared to the control group (P<0.0001) and a significantly increase in the proportion of positive neutrophils (VacR, 29.7%; PBS, 13.1%) and the mean fluorescent index (VacR, 217.4; PBS, 152.6; P<0.01) in the VacR group. The results suggest that VacR is a valuable vaccine candidate against S. aureus infections, and merits further field trials and experimental challenges.

Authors : Pujato N, Camussone C M, Renna M S, Perrig M S, Morein B, Calvinho L F, Marcipar I S,

(5) Changes in the content of immunostimulating components of colostrum obtained from dairy cows at different level of production.[TOP]

Pubmed ID :29604073
Publication Date : //
Recent data indicate that almost 60 percent of colostrum samples do not have enough antibodies to provide adequate protection for the calf. Thus, the aim of this study was to determine the influence of the level of production of cows on the content of immunostimulating components of protein fraction of colostrum. The observations were conducted on 2 farms that keep dairy cows. One of the farms was selected to represent typical performance level for the mass population of dairy cows in Poland, about 5500 kg of milk per cow per year (LI) and the second one was characterized by a higher level of production, about 8000 kg (HI).

Authors : Wąsowska Edyta, Puppel Kamila,

(6) Allantoin and hydantoin as new protein aggregation suppressors.[TOP]

Pubmed ID :29505874
Publication Date : //
Allantoin is widely used in pharmaceutical and cosmetic products, and is composed of a hydantoin ring and a ureido group. Recent reports showed that allantoin suppresses thermal aggregation of hen egg white lysozyme (LYZ). However, structural insight into the properties of allantoin on protein aggregation and whether allantoin controls the aggregation of other proteins under different stress conditions remain unclear. Here we studied the structural properties of allantoin in terms of its effects on protein aggregation by comparing allantoin with urea and hydantoin. Furthermore, we analyzed the effects of allantoin and its derivatives on the aggregation of LYZ, carbonic anhydrase from bovine erythrocytes (BCA), albumin from chicken egg white (OVA), and immunoglobulin G (IgG) by various stresses in comparison with arginine. These four proteins are widely different in charged state and molecular size. Allantoin suppressed the aggregation and inactivation of LYZ comparing to arginine without affecting the melting temperature of proteins, and was responsible for the slightly improved formation of soluble oligomers and insoluble aggregates of IgG with thermal and acidic stresses. In contrast, hydantoin increased the solubility of aromatic amino acids more effectively than arginine and allantoin. The structural properties underlying the observed effects of allantoin as an aggregation suppressor include hydrophobic interactions between hydantoin moiety and aromatic ring on the surface of proteins, which is reflected on the difference between allantoin and arginine. These results show that the backbone of hydantoin ring may be a new category of additives for development of small aggregation suppressors.

Authors : Nishinami Suguru, Yoshizawa Shunsuke, Arakawa Tsutomu, Shiraki Kentaro,

(7) The antibody response in the bovine mammary gland is influenced by the adjuvant and the site of subcutaneous vaccination.[TOP]

Pubmed ID :29490692
Publication Date : //
Intramammary infections in cattle resulting in mastitis have detrimental effects on cows' well-being, lifespan and milk production. In the host defense against S. aureus mastitis antibodies are thought to play an important role. To explore potential ways to increase antibody titers in the bovine mammary gland the effects of various adjuvants on the magnitude, isotype, and neutralizing capacity of antibodies produced following subcutaneous vaccine administration at different immunization sites were analyzed. In this study, α-toxoid was used as a model antigen and formulated in three different alum-based adjuvants: Alum-Saponin, Alum-Oil, and Alum-Saponin-Oil. Vaccines were administered near the suspensory ligament of the udder or in the lateral triangular area of the neck. At both immunization sites, immunization with α-toxoid in Alum-Saponin-Oil resulted in higher specific antibody titers in milk and serum as compared with Alum-Oil and Alum-Saponin, without favoring an IgG1, IgG2, or IgA response. Furthermore, the neutralizing capacity of milk serum and serum following immunization near the udder and in the neck was higher when Alum-Saponin-Oil was used as adjuvant compared with Alum-Oil and Alum-Saponin. Prime immunizations near the udder effectively increased both antibody isotype titers and neutralization titers, while prime plus boost immunizations were required to induce similar effects following immunization in the neck. Results indicate that subcutaneous administration of an Alum-Saponin-Oil based vaccine near the udder could be further explored for the development of a one-shot vaccination strategy to efficiently increase intramammary antibody responses.

Authors : Boerhout Eveline M, Koets Ad P, Mols-Vorstermans Tanja G T, Nuijten Piet J M, Hoeijmakers Mathieu J H, Rutten Victor P M G, Bijlsma Jetta J E,

(8) Pharmacological characterisation of CR6086, a potent prostaglandin E receptor 4 antagonist, as a new potential disease-modifying anti-rheumatic drug.[TOP]

Pubmed ID :29490676
Publication Date : //
Prostaglandin E (PGE) acts via its EP4 receptor as a cytokine amplifier (e.g., interleukin [IL]-6) and induces the differentiation and expansion of inflammatory T-helper (Th) lymphocytes. These mechanisms play a key role in the onset and progression of rheumatoid arthritis (RA). We present the pharmacological characterisation of CR6086, a novel EP4 receptor antagonist, and provide evidence for its potential as a disease-modifying anti-rheumatic drug (DMARD).

Authors : Caselli Gianfranco, Bonazzi Albino, Lanza Marco, Ferrari Flora, Maggioni Daniele, Ferioli Cristian, Giambelli Roberto, Comi Eleonora, Zerbi Silvia, Perrella Marco, Letari Ornella, Di Luccio Elena, Colovic Milena, Persiani Stefano, Zanelli Tiziano, Mennuni Laura, Piepoli Tiziana, Rovati Lucio Claudio,

(9) Generation of murine monoclonal antibodies with specificity against conventional camelid IgG1 and heavy-chain only IgG2/3.[TOP]

Pubmed ID :29475500
Publication Date : //
Camelids possess antibodies with a conventional four-chain structure consisting of two heavy and two light chains (of subclass IgG1) but further they also generate heavy-chain only antibodies (of subclass IgG2 and 3) which are fully functional in antigen binding. In this study subclass-specific murine monoclonal antibodies specific to conventional camelid IgG1 and heavy-chain only IgG2/3 were generated and validated for the use as potent secondary detection reagents. The monoclonal antibodies are able to differentiate between all camelid IgGs, conventional four-chain camelid antibodies (of subclass IgG1) and exclusively heavy chain-only antibodies (of subclasses IgG2 and IgG3). Further these antibodies were used to detect specific immune responses after vaccination of Camelids against bovine corona- and rotavirus strains and different E.coli and Clostridia - antigens and to identify Erysipelothrix rhusiopathiae infected animals within a herd. The described antibodies are suitable as new secondary agents for the detection of different camelid subclasses and the validation of camelid immune reactions.

Authors : Holzlöhner Pamela, Butze Monique, Maier Natalia, Hebel Nicole, Schliebs Erik, Micheel Burkhard, Füner Jonas, Heidicke Gabriele, Hanack Katja,

(10) Immune parameters to p67C antigen adjuvanted with ISA206VG correlate with protection against East Coast fever.[TOP]

Pubmed ID :29429808
Publication Date : //
East Coast fever (ECF) is a lymphoproliferative disease caused by the tick-transmitted protozoan parasite Theileria parva. ECF is one of the most serious cattle tick-borne diseases in Sub-Saharan Africa. We have previously demonstrated that three doses of the C-terminal part of the sporozoite protein p67 (p67C) adjuvanted with ISA206VG confers partial protection against ECF at a herd level. We have tested the efficacy of two doses of this experimental vaccine, as reducing the vaccination regimen would facilitate its deployment in the field. We reconfirm that three antigen doses gave a significant level of protection to severe disease (46%, ECF score < 6) when compared with the control group, while two doses did not (23%). Animals receiving three doses of p67C developed higher antibody titers and CD4 T-cell proliferation indices, than those which received two doses. A new panel of immune parameters were tested in order to identify factors correlating with protection: CD4 proliferation index, total IgG, IgG1, IgG2 and IgM half maximal titers and neutralization capacity of the sera with and without complement. We show that some of the cellular and humoral immune responses provide preliminary correlates of protection.

Authors : Lacasta Anna, Mwalimu Stephen, Kibwana Elisabeth, Saya Rosemary, Awino Elias, Njoroge Thomas, Poole Jane, Ndiwa Nicholas, Pelle Roger, Nene Vishvanath, Steinaa Lucilla,