Aim: To generate and identify monoclonal antibodies (mAb) against homo sapiens hydroxysteroid 11-beta dehydrogenase 1 (HSD11B1).
Methods: Normal human liver tissues were homogenized, and mitochondria were isolated by differential centrifugation. The total mitochondrial proteins were used to immunize BALB/c mice to generate mAb by hybridoma technique. The mAb were characterized by ELISA, Western blot and immunohistochemistry. The antibody specificity was identified by immunoprecipitation (IP), and confirmed by Uni-ZAP expression library screening.
Results: One hybridoma CBF245 secreting specific mAb against HSD11B1 was established. The Ig subclass of this mAb was IgG1, and it could be used in ELISA, Western blot, immunohistochemistry. Our data showed that the antigen recognized by CBF245 mAb was localized in the hepatocyte cytoplasm, with molecular weight of M(r) 35 000 in the mitochondria of human liver tissue. The CBF245 mAb was further confirmed by immunoscreening of Uni-ZAP XR liver cDNA expression library.
Conclusion: A hybridoma cell line stably secreting specific mAb against HSD11B1 is established and characterized. This mAb reacted with HSD11B1 in ELISA, Western blot, immunohistochemistry assay, IP, and would be very useful for the HSD11B1 studies.
Download full-text PDF |
Source |
|---|
Publication Analysis
Top Keywords
Similar Publications
Automated, Quantitative Capillary Western Blots to Analyze Host Cell Proteins in COVID-19 Vaccine Produced in Vero Cell Line.
Vaccines (Basel)
December 2024
Analytical Research & Development, Merck & Co., Inc., Rahway, NJ 07065, USA.
Background/objectives: Host cell protein (HCP) content is a major attribute for biological and vaccine products that must be extensively characterized prior to product licensure. Enzyme Linked Immunosorbent Assay (ELISA) and Mass Spectrometry (MS) are conventional methods for quantitative host cell protein analysis in biologic and vaccine products. Both techniques are usually very tedious, labor-intensive, and challenging to transfer to other laboratories.
View Article and Find Full Text PDFAerosol Inhalation of Luteolin-7-O-Glucuronide Exerts Anti-Inflammatory Effects by Inhibiting NLRP3 Inflammasome Activation.
Pharmaceuticals (Basel)
December 2024
Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, No. 16, Nanxiao Street, Dongzhimen, Dongcheng District, Beijing 100700, China.
Luteolin-7-O-glucuronide (L7Gn) is a flavonoid isolated from numerous traditional Chinese herbal medicines that exerts anti-inflammatory effects. Previous research has revealed that aerosol inhalation is the most straightforward way of administration for the delivery of respiratory agents. Thus far, the impact of aerosol inhalation of L7Gn on lung inflammation and the underlying mechanisms remain unknown.
View Article and Find Full Text PDFEvaluating the Diagnostic Efficacy of Using Pooled Samples for Chronic Wasting Disease Testing and Surveillance.
Pathogens
December 2024
Department of Veterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA 50011, USA.
Disease monitoring informs the opportunities for intervention by natural resource agencies tasked with managing chronic wasting disease (CWD) in wild cervids. However, allocating funds toward testing can reduce those available for education, outreach, and disease reduction. Implementation of more efficient testing strategies can help meet both an expanding need by resource managers and a burgeoning demand from the hunting public in North America.
View Article and Find Full Text PDFStructural and Serological Characterization of Yet Another New O Antigen, O86, in Clinical Strains.
Int J Mol Sci
December 2024
N. D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, 119991 Moscow, Russia.
Bacteria from the genus are facultative human pathogens, primarily attacking the urinary tract and wounds. A total of 85 O serogroups have been identified so far among these bacilli. Bprz 86 was isolated from the fistula of a patient in Łódź, Poland.
View Article and Find Full Text PDFRemote Ischemic Post-Conditioning (RIC) Mediates Anti-Inflammatory Signaling via Myeloid AMPKα1 in Murine Traumatic Optic Neuropathy (TON).
Int J Mol Sci
December 2024
Department of Translational Neuroscience, Barrow Neurological Institute, St Joseph's Hospital and Medical Center (SJHMC), Phoenix, AZ 85013, USA.
Traumatic optic neuropathy (TON) has been regarded a vision-threatening condition caused by either ocular or blunt/penetrating head trauma, which is characterized by direct or indirect TON. Injury happens during sports, vehicle accidents and mainly in military war and combat exposure. Earlier, we have demonstrated that remote ischemic post-conditioning (RIC) therapy is protective in TON, and here we report that AMPKα1 activation is crucial.
View Article and Find Full Text PDFWant 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!