Aim: To prepare rabbit polyclonal antibodies (pAb) and mouse monoclonal antibodies (mAbs) against human clusterin(CLU) and characterize these antibodies' properties.
Methods: CLU fragment was amplified from human liver cDNA library, and recombinant expression vectors pGEX-4T-1-CLU and PET-32a-CLU were constructed. GST-CLU fusion protein was expressed in E.coli and then used as the immunogen. Properties of antiserum against human CLU were identified by ELISA, Western blot, and the mAbs against human CLU was characterized by Western blot, indirect immunofluorescent staining and immunohistochemistry staining.
Results: The GST-CLU fusion protein was highly expressed with a molecular weight of M(r)54,000. Western blot analysis proved that the rabbit pAb could specifically recognize 52,000 and 58,000 proteins in human liver total protein. All of the nine established mAbs recognized recombinant human CLU protein, two of which specifically bound to proteins in the cytoplasm of HepG2 cells and four of which specifically bound to proteins in the cytoplasm of adult liver tissue.
Conclusion: pAb and mAbs against human CLU were successfully prepared, which will provide efficient tools for functional studies of CLU expressed in human tumors.
Download full-text PDF |
Source |
|---|
Publication Analysis
Top Keywords
Similar Publications
Targeting immune checkpoints on myeloid cells: current status and future directions.
Cancer Immunol Immunother
January 2025
Department of General Surgery, Shengjing Hospital of China Medical University, ShenyangLiaoning Province, 110004, China.
Myeloid cells accumulate extensively in most tumors and play a critical role in immunosuppression of the tumor microenvironment (TME). Like T cells, myeloid cells also express immune checkpoint molecules, which induce the immunosuppressive phenotype of these cells. In this review, we summarize the tumor-promoting function and immune checkpoint expression of four types of myeloid cells: macrophages, neutrophils, dendritic cells, and myeloid-derived suppressor cells, which are the main components of the TME.
View Article and Find Full Text PDFSingle-cell profiling uncovers synovial fibroblast subpopulations associated with chondrocyte injury in osteoarthritis.
Front Endocrinol (Lausanne)
December 2024
Spinal Surgery, Zhejiang Chinese Medical University Affiliated Wenzhou Hospital of Integrated Traditional Chinese and Western Medicine, Wenzhou, Zhejiang, China.
Article Synopsis
- This study focuses on understanding how synovial cells and chondrocytes interact in osteoarthritis (OA), aiming to clarify their roles in OA development.
- Using single-cell sequencing, researchers analyzed the characteristics of synovial fibroblasts and their interactions with chondrocytes, revealing significant correlations between these cell types in both damaged and healthy cartilage.
- The findings indicated that some genes in synovial fibroblasts promote chondrocyte health, while others contribute to chondrocyte degradation and inflammation, highlighting a complex balance that affects OA progression.
A non-invasive model for diagnosis of primary Sjogren's disease based on salivary biomarkers, serum autoantibodies, and Schirmer's test.
Arthritis Res Ther
December 2024
Department of Rheumatology and Clinical Immunology, School of Medicine, The First Affiliated Hospital of Xiamen University, Xiamen University, 55 Zhenhai Road, Xiamen, XM, 361000, China.
Article Synopsis
- The study focuses on identifying salivary proteins as potential biomarkers for primary Sjögren's disease (pSjD) to create a non-invasive prediction model.
- Liquid chromatography-tandem mass spectrometry was used on saliva samples to find proteins associated with immune processes, highlighting specific proteins such as CFB, CLU, and NE as independent predictors of pSjD.
- The proposed model combining these biomarkers with serum autoantibodies and Schirmer's test showed high accuracy, with a sensitivity of 84.85% and specificity of 92.45%, suggesting it could serve as a valuable non-invasive tool for pSjD classification.
Glycosylation Pattern of Serum Clusterin in Psoriatic Arthritis and Rheumatoid Arthritis-The Search for New Diagnostic Glycomarkers.
Int J Mol Sci
December 2024
Department of Rheumatology and Internal Medicine, Wroclaw Medical University, Borowska Street 213, 50-556 Wroclaw, Poland.
Psoriatic arthritis (PsA) and rheumatoid arthritis (RA) are connective tissue autoimmune diseases. The present study aimed to check whether serum clusterin (CLU) concentration and its glycosylation pattern may be markers differentiating these diseases-blood sera of patients with PsA (n = 37), RA (n = 34), and healthy subjects (control, n = 21) were examined. CLU concentration was measured using the ELISA test.
View Article and Find Full Text PDFIGHG4: innovative diagnostic biomarkers for iron overload in β-thalassemia patients.
Hematology
December 2025
Department of Pediatrics, The First Affiliated Hospital of Guangxi Medical University, Nanning, People's Republic of China.
Objectives: This study aims to investigate the serotransferrin (TF), complement C1s subcomponent (C1S), immunoglobulin heavy constant gamma 4 (IGHG4), hemoglobin subunit alpha (HBA1), and clusterin (CLU) contents in β-thalassemia patients, and explores their physiological role as potential non-invasive bioindicators for disease diagnosis and iron overload.
Methods: A total of 62 children with β-thalassemia were recruited and categorized by genotype, along with 17 healthy pediatric volunteers for analysis. The circulating ferritin content was evaluated, and plasma levels of TF, C1S, IGHG4, HBA1, and CLU were assessed using ELISA.
Want AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!