ThomasTu – ICE-HBV

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HBV Research Protocols

A modified kit-based HBV protein-free DNA extraction from liver tissues and cell cultures for HBV cccDNA Southern blot and qPCR
- cccDNA quantification from total DNA preparation by qPCR with cccDNA-selective primers is challenging because of the coexisting HBV DNA forms (replicative intermediates, protein-free ... [Read more]
21 September, 2022
A sensitive and rapid Southern blot assay based on branched DNA technology for the detection of HBV DNA in cell culture and liver tissue samples
- Sothern blot is considered the “gold standard” for cccDNA detection because it can separate the cccDNA from protein-free relaxed circular DNA (pf-rcDNA, aka deproteinated rcDNA (DP-rc... [Read more]
21 September, 2022
An Optimized Ex Vivo Flurospot assay to identify multi-functional HBV-specific T cells in Chronic Hepatitis B patients

During a traditional FluroSpot assay, low HBV Specific T cell frequencies have hindered effective ex vivo analysis. We overcame this obstacle to measure ex vivo T cell responses in CHB patients, ... [Read more]

07 March, 2022
Fluorescent-bait labelling for the ex vivo detection of HBV antigen-specific B cells

Fluorescently conjugated antigen-bait systems have been extensively used to identify antigen-specific B cells and probe humoral immunity across different settings. Using this principle, HBV antig... [Read more]

30 April, 2021
Tetramer enrichment for HBV-specific CD8+ T cells

The frequency of HBV-specific CD8+ T cells can be very low, depending on the specificity. Therefore, they cannot always be detected by conventional ex vivo tetramer staining using 10[Read more]

03 July, 2020
A fluorescent in situ hybridization (FISH) assay for detection of HBV DNA in cell culture models
- Although a plethora of knowledge on the molecular life cycle of Hepatitis B Virus has been gained by utilizing classical methods, they can only inform on the average level of the test... [Read more]
19 June, 2020
HBV Ribonuclease H assay resolved by denaturing PAGE
- This assay characterizes the products of HBV RNase H activity on a DNA:RNA heteroduplex.
- Products are resolved by denaturing PAGE to assess specificity of cleavage and eff... [Read more]
06 June, 2019
HBV Ribonuclease H FRET Assay
- This assay detects the activity of HBV RNaseH using fluorescence resonance energy transfer
- The ribonuclease H (RNaseH) substrate is an RNA DNA heteroduplex in which an RNA... [Read more]
06 June, 2019
A Southern Blot Assay for Detection of HBV cccDNA from Cell Cultures
- As an essential component of HBV life cycle, the viral covalently closed circular DNA (cccDNA) is synthesized and maintained at low copy numbers in the nucleus of infected hepatocytes... [Read more]
03 June, 2019
Modeling HBV infection and therapy in immunodeficient NOD-Rag1-/-IL2RgammaC-null (NRG) fumarylacetoacetate hydrolase (FAH) knockout mice with human chimeric liver
- Besides human, only a few species have been reported to be permissive for Hepatitis B virus (HBV) infection(1-3). Human liver chimeric mouse models have been developed to overcome the... [Read more]
25 May, 2019