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#28724556   2017/07/20 Save this To Up

Performance assessment of a Trypanosoma cruzi chimeric antigen in multiplex liquid microarray assays.

Diagnosing chronic Chagas disease (CD) requires antibody--antigen detection methods, traditionally based on enzymatic assay techniques whose performance depend on the type and quality of antigen used. Previously, 4 recombinant chimeric proteins from Instituto de Biologia Molecular do Paraná (IBMP-8.1 to -8.4) comprising immuno-dominant regions of diverse Trypanosoma cruzi antigens showed excellent diagnostic performance in enzyme-linked immunosorbent assays. Considering that next-generation platforms offer improved CD diagnostic accuracy with different T. cruzi-specific recombinant antigens, we assessed the performance of these chimeras in liquid microarrays (LMAs). The chimeric proteins were expressed in Escherichia coli and purified by chromatography. Sera from 653 chagasic and 680 healthy individuals were used to assess the performance of these chimeras in detecting specific anti-T. cruzi antibodies. Accuracies ranged from 98.1--99.3%, and diagnostic odds ratio values were 3,548 for IBMP-8.3, 4,826 for IBMP-8.1, 7,882 for IBMP-8.2, and 25,000 for IBMP-8.4. A separate sera bank (851 samples) was employed to assess cross-reactivity with other tropical diseases. Leishmania spp., a pathogen with high genome sequence similar to T. cruzi, showed cross-reactivity rates ranging from 0--2.17%. Inconclusive results were negligible (0--0.71%). Bland--Altman and Deming regression analysis based on 200 randomly selected CD-positive and -negative samples demonstrated interchangeability with respect to CD diagnostic performance in both singleplex and multiplex assays. Our results suggested that these chimeras can potentially replace antigens currently used in commercially available assay kits. Moreover, the use of a multiplex platform, like LMA assays employing 2 or more IBMP antigens, would abrogate the need for 2 different testing techniques when diagnosing CD.

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HIV 1 intergase antigen. Caspase-3 Substrate DEVD- Caspase-3 Substrate DEVD- Caspase-3 Substrate DEVD- Caspase-3 Substrate DEVD- Caspase-3 Inhibitor Z-DEV Caspase-3 Inhibitor Z-DEV Caspase-Family Inhibitor Caspase-Family Inhibitor Caspase-6 Inhibitor Z-VEI Caspase-6 Inhibitor Z-VEI Caspase-1 Inhibitor Z-YVA

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#28719742   2017/07/18 Save this To Up

Integrated smartphone-app-chip system for on-site ppb-level colorimetric quantitation of aflatoxins.

We demonstrate herein an integrated, smartphone-app-chip (SPAC) system for on-site quantitation of food toxins, as demonstrated with aflatoxin B1 (AFB1), at parts-per-billion (ppb) level in food products. The detection is based on an indirect competitive immunoassay fabricated on a transparent plastic chip with the assistance of a microfluidic channel plate. A 3D-printed optical accessory attached to a smartphone is adapted to align the assay chip and to provide uniform illumination for imaging, with which high-quality images of the assay chip are captured by the smartphone camera and directly processed using a custom-developed Android app. The performance of this smartphone-based detection system was tested using both spiked and moldy corn samples; consistent results with conventional ELISA kits were obtained. The achieved detection limit (3±1 μg/kg, equivalent to ppb) and dynamic response range (0.5 to 250 μg/kg) meet the requested testing standards set by authorities worldwide. We envision that the integrated SPAC system promises to be a simple and accurate method of food toxin quantitation, bringing much benefit for rapid on-site screening.

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Amplite™ Colorimetric A Amplite™ Colorimetric A Amplite™ Fluorimetric F Amplite™ Colorimetric U Amplite™ Colorimetric C Amplite™ Colorimetric B Amplite™ Colorimetric M Cellufine Formyl , 50 ml Cellufine Formyl Media Cellufine Formyl , 500 ml Cellufine Formyl Media Cellufine Formyl Media

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#28719352   2017/07/18 Save this To Up

Determination of IL-6, TNF-α and VEGF levels in the serums of patients with colorectal cancer.

Cytokines are multifunctional polypeptides synthesized by different body cells. They have clinical significance in terms of disease diagnosis, treatment and prevention. Cytokines TNF-α and IL-6 play an important role in the growth and differentiation of cells.Vascular Endothelial Growth Factor (VEGF) is excessively produced in epithelial, mesenchymal, and particularly in tumor cells. Studies have shown that the increased serum concentrations of IL-6, TNF-α, VEGF are strongly associated with colorectal cancer  and directly with the clinical stage of the disease. This can be used to diagnose cancer and to identify patients with a bad prognosis who can avail themselves of a more aggressive treatment. The present study investigated the role of cytokines in the development of cancer by comparing preoperative serum cytokine levels of patients suffering from colorectal cancer with those of the healthy control group. The prognostic significance of the data obtained has also been evaluated. For this purpose, IL-6, TNF-α and VEGF levels in 60 serums, 30 preoperatively taken from patients with colorectal cancer and 30 from a healthy control group at Çanakkale Onsekiz Mart University General Surgery Clinic, were determined by ELISA kits. The statistical analyses of the obtained data were evaluated on SPSS, a statistical package program. In this study, no significant difference was obtained between the mean scores concerning the IL-6 and VEGF serums of the colorectal cancer and healthy group (p>.05). But a statistically significant decrease was observed in the TNF-α serum level of the colorectal cancer group in comparison with the control group (p= .016; p < .05).

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CELLKINES Natural Human I Human Interleukin-4 IL-4 Human Interleukin-6 IL-6 Human Interleukin-7 IL-7 Human Interleukin-2 IL-2 Human Interleukin-16 IL-1 Human Interleukin-33 IL-3 Human Interleukin-17E (IL Human Interleukin-32 alph Human Interleukin-17F IL- Human Interleukin-17AF He Human Epstein-Barr Virus

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#28716207   2017/07/18 Save this To Up

Influence of true within-herd prevalence of small ruminant lentivirus infection in goats on agreement between serological immunoenzymatic tests.

The study was conducted to evaluate influence of the true within-herd prevalence of small ruminant lentivirus (SRLV) infection on agreement beyond chance between three different types of commercial serological ELISAs. Blood samples were collected from 865 goats from 12 dairy goat herds. Serum samples were tested using three commercial ELISA kits: whole-virus indirect ELISA (wELISA), indirect ELISA based on recombined TM and CA antigens (TM/CA-ELISA), and competitive-inhibition ELISA based on SU antigen (SU-ELISA). Herds were classed into three prevalence strata of high (>50%), moderate (10-50%) and low (<10%) true within-herd prevalence of SRLV infection. The latter was estimated on the basis of results of wELISA adjusted by its sensitivity and specificity. Agreement beyond chance between the three ELISAs was assessed at two levels. First, the general agreement was determined using two coefficients corrected for chance-agreement: Cohen's kappa and Gwet's AC1. Then, agreement between tests was evaluated using Gwet's AC1 separately in the three prevalence strata and compared between them by computing 95% confidence intervals for differences with a Bonferroni correction for multiple comparisons. The general agreement between the three tests was very good: wELISA and TM/CA-ELISA - Cohen's kappa of 81.8% (CI 95%: 77.9% to 85.7%), Gwet's AC1 of 82.7% (CI 95%: 79.0% to 86.4%); wELISA and SU-ELISA - Cohen's kappa of 83.2% (CI 95%: 79.4% to 86.9%), Gwet's AC1 of 83.9% (CI 95%: 80.4% to 87.5%); TM/CA-ELISA and SU-ELISA - Cohen's kappa of 86.0% (CI 95%: 82.6% to 89.5%), Gwet's AC1 of 86.9% (CI 95%: 83.6% to 90.1%). However, agreement between ELISAs was significantly related to the within-herd true prevalence - it was significantly lower (although still high) when within-herd true prevalence was moderate (Gwet's AC1 between 67.2% and 78.7%), whereas remained very high, when true within-herd prevalence was either >50% (Gwet's AC1 between 91.9% and 98.8%) or <10% (Gwet's AC1 between 94.7% and 98.4%). Concluding, the three different commercial ELISAs for SRLV infection in goats available on the market yield highly consistent results. However, their agreement is affected by the true within-herd prevalence in a tested population, and the worse (although still high) agreement should be expected, when the percentage of infected goats is moderate.

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#28713178   2017/07/17 Save this To Up

Glucose transporter-4 in white blood cells of young and old sled dogs: a model for human biomarker development.

The insulin responsive glucose transporter, GLUT4 is found predominantly in muscle and adipose cells. Maratou and others (2007) reported that there is GLUT4 in white blood cells (WBC) collected from human subjects in response to insulin activation. This study was designed to validate the presence of GLUT4 in white blood cells of sled dogs and furthermore to investigate whether changes in levels of the GLUT4 protein might be associated with aging. Additionally, we examined the blood insulin concentration of two populations of dogs, young and old, before and after a meal to observe their insulin response. It is documented in skeletal muscle that GLUT4 expression is increased as a result of conditioning, making sled dogs an excellent model in the circumpolar north for studying the effects of exercise, nutrition and diabetes (Felsburg 2002; Kararli 2006). Blood was withdrawn from 11 healthy sled dogs: 6 young (1-5 years) and physically fit, conditioned for racing and 5 old (7-13 years), retired from racing. The insulin response was determined using blood plasma and ELISA. The buffy coat (containing WBC) was collected with a glass pipette after centrifugation and washed and suspended in 1x phosphate buffer. GLUT4 was measured using ELISA kits (USCN Life Sciences). The results validate that GLUT4 is present in white blood cells in sled dogs. Age had no significant effect in the concentration of GLUT4 between the populations of old and young dogs. A significant difference in insulin levels pre and post meal in young (0.13 ± 0.03 ng/mL (pre), 0.22 ± 0.04 ng/mL (post), p < 0.05) and old (0.13 ± 0.02 ng/mL (pre), 0.22 ± 0.03 ng/mL (post), p < 0.05) dogs was observed, displaying the typical postprandial insulin spike. No significant difference was found in insulin concentration comparing old versus young dogs. Our data shows that white blood cells in young (40.4 ± 2.4 ng/mL) and old (35.3 ± 8.8 ng/mL) sled dogs have quantifiable but non-significant different GLUT4 levels (p > 0.05). Detecting GLUT4 via an ELISA in white blood cells, opens up minimally invasive avenues for studying the underlying molecular mechanisms associated with insulin resistance in more complex, dynamic and physiological systems. This project was the first step in developing a protocol for this simple, technique with a potential clinical application for diagnosing insulin resistance.

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#28713974   2017/07/17 Save this To Up

miRNA-125b regulates apoptosis of human non-small cell lung cancer via the PI3K/Akt/GSK3β signaling pathway.

The present investigation demonstrated that regulation of microRNA (miR)-125b affected the apoptosis of human non-small cell lung cancer (NSCLC) through targeting of the PI3K/Akt and Wnt/β-catenin signaling pathways. The expression of miR-125b was assessed in patients with NSCLC, which demonstrated that miR-125b expression in NSCLC tissue was higher than that in para-carcinoma tissue. Furthermore, survival analysis of patients with NSCLC over 3 years indicated that the overall survival (OS) and disease-free survival (DFS) rates of patients with low miR-125b expression were higher than those of patients with high miR-125b expression. Proliferation and apoptosis assays were subsequently conducted in the human NSCLC cell line A549 using MTT assay and Annexin V-FITC/PI kits, respectively. Caspase-3 activity ELISA and western blot analysis were also used to assess caspase-3 activity and the protein expression of Bax, Akt, phosphorylated (p)-Akt, p-GSK3β, Wnt and β-catenin. It was observed that downregulation of miR-125b inhibited the proliferation and induced the apoptosis of A549 cells. Downregulation of miR-125b also suppressed the protein expression of p-Akt, Wnt and β-catenin, and increased caspase-3 activity and Bax protein expression in A549 cells. In addition, downregulation of miR-125b combined with the PI3K inhibitor LY294002 enhanced cell growth inhibition, suppression of p-GSK3β, Wnt and β-catenin protein expression and promotion of caspase-3 activity in A549 cells. These results revealed that the downregulation of miR-125b regulates apoptosis in human NSCLC through the suppression of the PI3K/Akt/GSK3β and Wnt/β-catenin signaling pathways.

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#28709448   2017/07/15 Save this To Up

No added diagnostic value of non-phosphorylated tau fraction (p-taurel) in CSF as a biomarker for differential dementia diagnosis.

The Alzheimer's disease (AD) cerebrospinal fluid (CSF) biomarkers Aβ1-42, t-tau, and p-tau181 overlap with other diseases. New tau modifications or epitopes, such as the non-phosphorylated tau fraction (p-taurel), may improve differential dementia diagnosis. The goal of this study is to investigate if p-taurel can improve the diagnostic performance of the AD CSF biomarker panel for differential dementia diagnosis.

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#28700471   2017/07/12 Save this To Up

Plasma cytokines can help to identify the development of severe acute pancreatitis on admission.

Severe acute pancreatitis (AP) is associated with high morbidity and mortality. Early severity stratification remains a challenging issue to overcome to improve outcomes. We aim to find novel plasma cytokines for the early identification of severe AP according to the revised Atlanta criteria.In this prospective observational study, 30 cytokines, screened semiquantitatively with a human multicytokine array, were submitted to quantitative determination using either microparticle-based multiplex immunoassays analyzed on a Luminex 100 platform or enzyme-linked immunosorbent assay kits. The cytokine profiles of patients and the discriminative value of cytokines for severe AP were analyzed.Plasma samples of 70 patients with AP (20 mild, 30 moderately severe, and 20 severe) were selected in this study if they were admitted within 48 hours of the onset of symptoms. Plasma from healthy volunteers was collected as the healthy control. Growth differentiation factor-15 (GDF-15) and pentraxin 3 (PTX3) on admission were independent prognostic markers for the development of severe AP and had higher discriminative powers than conventional markers (GDF-15 vs hematocrit, P = .003; GDF-15 vs C-reactive protein, P = .037; GDF-15 vs creatinine, P = .048; GDF-15 vs Acute Physiology and Chronic Health Evaluation II, P = .007; PTX3 vs hematocrit, P = .006; PTX3 vs C-reactive protein, P = .047; PTX3 vs Acute Physiology and Chronic Health Evaluation II, P = .011; PTX3 vs Bedside Index for Severity in Acute Pancreatitis, P = .048).Plasma GDF-15 and PTX3 can help to identify the development of severe AP on admission. Future work should validate their accuracy in a larger, multicenter patient cohort.

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#28694931   2017/07/11 Save this To Up

Interleukin-18 polymorphism as an inflammatory index in metabolic syndrome: A preliminary study.

To assess circulatory levels of interleukin-18 (IL-18) and determine whether the presence of IL-18 promoter polymorphism influences metabolic syndrome phenotypes.

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#28692306   2017/07/10 Save this To Up

Development and validation of an electrochemiluminescent ELISA for quantitation of oral insulin tregopil in diabetes mellitus serum.

Tregopil, a novel PEGylated human insulin is in clinical development for oral delivery in diabetes treatment. The aim of the study was to develop and validate a sensitive and specific ELISA method for quantitating Tregopil in diabetes subjects on basal Glargine, since most commercially available insulin kits either do not detect Tregopil or show significant reactivity to Glargine.

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