Comparison of methods and characterization of small RNAs from plasma extracellular vesicles of HIV/HCV coinfected patients
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Comparison of methods and characterization of small RNAs from plasma extracellular vesicles of HIV/HCV coinfected patients
The hepatitis C virus (HCV) and human immunodeficiency virus (HIV) hijacks the host machine exosomal addition to the mechanism of infection and evasion of the immune system, modifying a small RNA (smRNA) cargo during infection. Our surface epitope tagged extracellular vesicles (EV) on plasma HIV / HCV-coinfected patients and their cargo smRNA profile, by comparing the different isolation procedures.
six EVS isolation procedures were compared: ultracentrifugation, and five polyethylene glycol based on a different method (commercial, combined with step purification column and two custom); and the two RNA commercial kit (phenol and non-phenol based) are used. High-throughput sequencing of smRNAs do.
Exosomal surface epitopes were analyzed by MACSPlex Exosome Kit. Four miRNAs shown the difference between protocol (HSA-miR-205-5p and HSA-let-7a / b / f-5p). Selection of RNA isolation kits impact on the detection of miRNAs and other smRNAs, where the phenol-based RNA isolation kits are made welcome. EVS enriched surface with HLA-DR / DP / DQ, CD81, and CD8. There are three liver-specific miRNAs expressed (let-7a-5p, miR-21-5p and HSA-miR-122-5p), thus, EVS cargo may reflect the evolution of liver disease.
Another smRNAs like piwi-interacting RNAs were also detected for the first time. Custom-based polyethylene glycol precipitation method combined with phenol-based RNA kit produces a higher number of smRNAs for EVs isolated from patients with HIV / HCV plasma.
micrographs was used to determine the crystal structure, western blotting was used to identify markers exosomal, and NanoSight used to characterize the nanoparticles. The relative expression levels of miRNAs which former is calculated using quantitative PCR and compared between patients with PC and CP. The expression levels of the two former miRNA-21 and miRNA-210 were significantly higher in patients with PC compared to patients with CP (both P <0.001).
However, no significant differences in serum levels are relatively free miR-21 and miR-210 were observed between the 2 groups of patients (both P> 0.05). ex-miRNA-21 and miRNA-210 was associated with tumor stage, as well as other factors. Ex diagnostic potential miRNA-21 and miRNA-210 levels were 83 and 85%, respectively. In addition, when the former Mirna and serum carbohydrate antigen 19-9 combined expression levels, accuracy is increased to 90%. This study identified that the serum former miRNAs, miRNA-21 and miRNA-210 may be of value as a potential biomarker and therapeutic target for the diagnosis and treatment of PC.
Comparison of methods and characterization of small RNAs from plasma extracellular vesicles of HIV/HCV coinfected patients
Evaluation exosome separation of human serum with fractional asymmetric flow field-flow-inlet frit and multiangle light scattering
Exosomes are extracellular vesicles that mediate communication between, the immune response, and tumor metastasis. However, the isolation of blood exosome complex because their size and density similar to blood lipoprotein.
Rat Complete SeraMir Exosome RNA Amplification and Profiling Kit for Media and Urine (contains cat# RA800TC-1 with 50ml ExoQuick-TC, RA805A-1 and RA812A-1 components)
Description: Lyophilized exosomes can be used as control standards for FACS, WB, ELISA and as calibration standards for quantitation of exosome-derived markers from biological samples.
Exosome isolation Solution - Serum/ Plasma (easy version of P101, 95% pure exosome)
Description: A sandwich quantitative ELISA assay kit for detection of Human Exosome Component 2 (EXOSC2) in samples from tissue homogenates, cell lysates or other biological fluids.
Description: A sandwich quantitative ELISA assay kit for detection of Human Exosome Component 2 (EXOSC2) in samples from tissue homogenates, cell lysates or other biological fluids.
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Human Exosome Component 7 (EXOSC7) in Tissue homogenates, cell lysates and other biological fluids.
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Human Exosome Component 7 (EXOSC7) in Tissue homogenates, cell lysates and other biological fluids.
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Human Exosome Component 7 (EXOSC7) in Tissue homogenates, cell lysates and other biological fluids.
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Human Exosome Component 7 (EXOSC7) in Tissue homogenates, cell lysates and other biological fluids.
Description: Enzyme-linked immunosorbent assay based on the Double-antibody Sandwich method for detection of Human Exosome Component 7 (EXOSC7) in samples from Tissue homogenates, cell lysates and other biological fluids with no significant corss-reactivity with analogues from other species.
Here, we employed programming field frit-inlet fractionation asymmetrical flow field-flow (FIAF4) coupled with light scattering multiangle (MALS) for effective separation of exosomes from the protein bound to the free and lipoproteins present in serum samples using the methods of pre-treatment is different, namely , a commercial kit exosome isolation, ultracentrifugation (UC), and a simple centrifugation followed by ultrafiltration (UF) .Sizes of exosomes eluted, as calculated by the MALS signal, approximated well with the results of dynamic light scattering batch of fraction was collected and the particle size of polystyrene
