RESUMO
Synaptotagmin-1 is a synaptic vesicle transmembrane protein that senses calcium influx via its tandem C2-domains, triggering synchronous neurotransmitter release. Disruption to SYT1 is associated with neurodevelopmental disorders, highlighting the importance of identifying high-quality research reagents to enhance understanding of Synaptotagmin-1 in health and disease. Here we have characterized thirteen Synaptotagmin-1 commercial antibodies for western blot, immunoprecipitation, immunofluorescence and flow cytometry using a standardized experimental protocol based on comparing read-outs in knockout cell lines and isogenic parental controls. These studies are part of a larger, collaborative initiative seeking to address antibody reproducibility issues by characterizing commercially available antibodies for human proteins and publishing the results openly as a resource for the scientific community. While use of antibodies and protocols vary between laboratories, we encourage readers to use this report as a guide to select the most appropriate antibodies for their specific needs.
Assuntos
Anticorpos , Western Blotting , Citometria de Fluxo , Imunofluorescência , Imunoprecipitação , Sinaptotagmina I , Sinaptotagmina I/imunologia , Sinaptotagmina I/metabolismo , Humanos , Citometria de Fluxo/métodos , Imunoprecipitação/métodos , Imunofluorescência/métodos , Anticorpos/imunologiaRESUMO
Background: Based on the Liver Imaging Data and Reporting System (LI-RADS) guidelines, Hepatocellular Carcinoma (HCC) can be diagnosed using imaging criteria in patients at risk of HCC. Objective: This study aimed to assess the diagnostic value of LI-RADS in high-risk patients with HCC. Material and Methods: This systematic review is conducted on international databases, including Google Scholar, Web of Science, PubMed, Embase, PROQUEST, and Cochrane Library, with appropriate keywords. Using the binomial distribution formula, the variance of each study was calculated, and all the data were analyzed using STATA version 16. The pooled sensitivity and specificity were determined using a random-effects meta-analysis approach. Also, we used the chi-squared test and I2 index to calculate heterogeneity among studies, and Funnel plots and Egger tests were used for evaluating publication bias. Results: The pooled sensitivity was estimated at 0.80 (95% CI 0.76-0.84). According to different types of Liver Imaging Reporting and Data Systems (LI-RADS), the highest pooled sensitivity was in version 2018 (0.83 (95% CI 0.79-0.87) (I2: 80.6%, P of chi 2 test for heterogeneity <0.001 and T2: 0.001). The pooled specificity was estimated as 0.89 (95% CI 0.87-0.92). According to different types of LI-RADS, the highest pooled specificity was in version 2014 (93.0 (95% CI 89.0-96.0) (I2: 81.7%, P of chi 2 test for heterogeneity <0.001 and T2: 0.001). Conclusion: LI-RADS can assist radiologists in achieving the required sensitivity and specificity in high-risk patients suspected to have HCC. Therefore, this strategy can serve as an appropriate tool for identifying HCC.
RESUMO
Vacuolar protein sorting-associated protein 35 is a subunit of the retromer complex, a vital constituent of the endosomal protein sorting pathway. The D620N mutation in the VPS35 gene has been reported to be linked to type 17 Parkinson's Disease progression, the exact molecular mechanism remains to be solved. The scientific community would benefit from the accessibility of validated and high-quality anti-hVPS35 antibodies. In this study, we characterized thirteen hVPS35 commercial antibodies for Western Blot, immunoprecipitation, and immunofluorescence using a standardized experimental protocol based on comparing read-outs in knockout cell lines and isogenic parental controls. We identified many high-performing antibodies and encourage readers to use this report as a guide to select the most appropriate antibody for their specific needs.
Assuntos
Anticorpos , Western Blotting , Imunofluorescência , Imunoprecipitação , Transporte ProteicoRESUMO
Tyrosine-protein kinase SYK, encoded by the SYK gene, is a non-receptor type protein kinase which mediates immune signal transduction through immunoreceptors. Tyrosine-protein kinase SYK expression has been associated with the development of various inflammatory diseases, cancer and neurodegenerative conditions. The reproducibility of tyrosine-protein kinase SYK research would help elucidate the mechanism in which it causes neuroinflammation as well as its potential as a novel target to treat Alzheimer's disease. This would be facilitated with the availability of high-quality tyrosine-protein kinase SYK. In this study, we characterized thirteen tyrosine-protein kinase SYK commercial antibodies for Western Blot, immunoprecipitation, and immunofluorescence using a standardized experimental protocol based on comparing read-outs in knockout cell lines and isogenic parental controls. We identified many high-performing antibodies and encourage readers to use this report as a guide to select the most appropriate antibody for their specific needs.
RESUMO
Moesin is a cytoskeletal adaptor protein, involved in the modification of the actin cytoskeleton, with relevance to Alzheimer's Disease. Well characterized anti-Moesin antibodies would benefit the scientific community. In this study, we have characterized ten Moesin commercial antibodies in Western Blot, immunoprecipitation, and immunofluorescence using a standardized experimental protocol based on comparing read-outs in knockout cell lines and isogenic parental controls. These studies are part of a larger, collaborative initiative seeking to address antibody reproducibility by characterizing commercially available antibodies for human proteins and publishing the results openly as a resource for the scientific community. While use of antibodies and protocols vary between laboratories, we encourage readers to use this report as a guide to select the most appropriate antibodies for their specific needs.
Assuntos
Anticorpos , Proteínas do Citoesqueleto , Humanos , Reprodutibilidade dos Testes , Proteínas do Citoesqueleto/metabolismo , Western Blotting , Imunoprecipitação , ImunofluorescênciaRESUMO
A member of the RNA-binding protein family, T-cell intracellular antigen-1 (TIA1) regulates mRNA translation and splicing as well as cellular stress by promoting stress granule formation. Variants of the TIA1 gene have implications in neurogenerative disorders including amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Reproducible research on TIA1 would be enhanced with the availability of high-quality anti-TIA1 antibodies. In this study, we characterized twelve TIA1 commercial antibodies for Western Blot, immunoprecipitation, and immunofluorescence using a standardized experimental protocol based on comparing read-outs in knockout cell lines and isogenic parental controls. We identified many high-performing antibodies and encourage readers to use this report as a guide to select the most appropriate antibody for their specific needs.
Assuntos
Proteínas de Ligação a RNA , Antígeno-1 Intracelular de Células T/genética , Western Blotting , Imunofluorescência , ImunoprecipitaçãoRESUMO
Rab1 is a highly conserved small GTPase that exists in humans as two isoforms: Rab1A and Rab1B, sharing 92% sequence identity. These proteins regulate vesicle trafficking between the endoplasmic reticulum (ER) and Golgi and within the Golgi stacks. Rab1A and Rab1B may be oncogenes, as they are frequently dysregulated in various human cancers. Moreover, they contribute to the progression of Parkinson's disease. The availability of high-quality antibodies specific for Rab1A or Rab1B is essential to understand the distinct functions of these Rab1 proteins in both health and diseaseand to enhance the reproducibility of research involving these proteins. In this study, we characterized seven antibodies targeting Rab1A and five antibodies targeting Rab1B for Western Blot, immunoprecipitation, and immunofluorescence using a standardized experimental protocol based on comparing read-outs in knockout cell lines and isogenic parental controls. These studies are part of a much larger, collaborative initiative seeking to address the antibody reproducibility issue by characterizing commercially available antibodies for human proteins and publishing the results openly as a valuable resource for the scientific community. While uses of antibodies and protocols vary between laboratories, we encourage readers to use this report as a guide to select the most appropriate antibodies for their specific needs.