RNASEH1 Antibody, Biotin conjugated
Description
Definition and Overview
The RNASEH1 Antibody, Biotin conjugated is a specialized immunological reagent designed to specifically target the ribonuclease H1 (RNASEH1) protein. Conjugated with biotin, this antibody enables detection via biotin-avidin systems, making it highly versatile for applications such as enzyme-linked immunosorbent assays (ELISA), immunoprecipitation (IP), and Western blotting (WB). RNASEH1 is an enzyme critical for resolving RNA-DNA hybrids, with roles in DNA replication, repair, and mitochondrial function .
Biotin Conjugation
Biotin is covalently linked to the antibody’s lysine residues, typically via NHS-biotin chemistry. This modification allows the antibody to bind streptavidin or avidin, enabling signal amplification in assays.
Antigen Specificity
The antibody is generated against a synthetic peptide corresponding to amino acids 1–75 of human RNASEH1 . This region is part of the N-terminal domain (NTD), which interacts with RNA-DNA hybrids and facilitates RNASEH1’s enzymatic activity .
Applications
The biotin-conjugated RNASEH1 antibody is optimized for:
ELISA
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Use Case: Quantitative detection of RNASEH1 in lysates or purified samples.
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Workflow: Capture antibody binds RNASEH1, followed by detection via biotin-streptavidin-HRP systems.
Immunoprecipitation (IP)
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Use Case: Isolation of RNASEH1-protein complexes for downstream analysis (e.g., mass spectrometry).
Western Blot (WB)
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Use Case: Qualitative detection of RNASEH1 in cell lysates or tissue extracts.
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Validation: Detected in HeLa, A549, HEK-293, and Jurkat cells .
Product-Specific Data
Functional Insights
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RNASEH1’s Role: Critical in resolving RNA-DNA hybrids during DNA repair and mitochondrial replication .
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Interactions: RNASEH1 binds proteins like NAT10 and DDX21 to enhance its activity, particularly in response to phosphorothioate oligonucleotides (PS-ASOs) .
Comparative Analysis of Biotin-Conjugated RNASEH1 Antibodies
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- RNASEH1 gene variants have been associated with susceptibility or protection against T1 Diabetes in Colombia. PMID: 29204916
- Research indicates that ribonuclease H1 is essential for mitochondrial DNA replication. PMID: 27402764
- Studies have shown that the catalytic domains of E. coli and human RNase H have nearly identical sequence preferences, which correlate with the efficiency of RNase H-recruiting antisense oligonucleotides. PMID: 29126318
- Evidence suggests that ribonuclease H1 (RNASEH1) plays a significant role in replication fork movement by resolving R-loops (RNA-DNA hybrids). Depletion of RNASEH1 leads to an accumulation of RNA-DNA hybrids, slowing of replication forks, and increased DNA damage. RNASEH1 appears to contribute to genome stability and preserves telomere integrity. PMID: 28717002
- RPA acts as a sensor of R loops and a regulator of RNaseH1, expanding the versatile role of RPA in suppressing genomic instability. PMID: 28257700
- RNaseH1 maintains regulated levels of telomeric RNA-DNA hybrids at ALT telomeres to trigger homologous recombination without compromising telomere integrity. PMID: 25330849
- Research has found that the 3' fragments of target pre-mRNA generated by ASO were almost completely degraded from their 5' ends by nuclear XRN2 after RNase H1-mediated cleavage. PMID: 26159921
- Altered RNaseH1 exhibits a reduced ability to remove RNA from RNA-DNA hybrids, leading to impaired mtDNA replication and adult-onset mitochondrial encephalomyopathy. PMID: 26094573
- RNase H1 and protein P32 are involved in mitochondrial pre-rRNA processing. PMID: 23990920
- Data implicate the H264 side chain in phosphodiester hydrolysis and product release, consistent with a proposed model where the RNAse H1 H264 side chain interacts with a divalent metal ion to support catalysis. PMID: 23078533
- Based on its nuclear magnetic resonance (NMR) nucleic acid structure, a boranophosphonate-modified, fully R(P) BH(3) DNA/RNA hybrid is predicted not to be a substrate for RNase H1. PMID: 21443203
- Observational study of gene-disease association. (HuGE Navigator) PMID: 20877624
- Site-directed mutagenesis studies have determined that the cysteine residues responsible for the redox-dependent activity of RNase H1 involve Cys(147) and Cys(148), producing an inactive enzyme conformation through disulfide bond formation. PMID: 12473655
- Human RNase H1 utilizes one tryptophan and two lysines to position the enzyme at the 3'-DNA/5'-RNA terminus of the heteroduplex substrate. PMID: 14506260
- In human cells, RNase H1 is responsible for most of the activity of DNA-like antisense drugs. PMID: 14960586
- Analysis of the catalytic site of human RNase H1 for heteroduplex substrate catalysis. PMID: 15205459
- Method for enhancing the human RNase H1 activity of chimeric antisense oligonucleotides. PMID: 17028157
- Research investigating the role of substrate structure in directing human RNase H1 activity, as well as the design of effective antisense oligodeoxyribonucleotides. PMID: 17028158
- Report of crystal structures of RNase H1 in complex with RNA/DNA hybrids. PMID: 17964265
- Characterization of full-length enzymes with defective hybrid binding domain indicates that this domain significantly enhances both the specific activity and processivity of RNase H1. PMID: 18337749
