HS3ST1 Antibody
Description
Structure and Mechanism of HS3ST1 Antibody
HS3ST1 antibodies are typically polyclonal or monoclonal, targeting specific epitopes of the HS3ST1 protein. Key structural features include:
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Immunogen: Most antibodies are raised against recombinant HS3ST1 protein or synthetic peptides, such as the C-terminal region (e.g., ab254718) or full-length sequences (e.g., HPA002237) .
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Host Species: Common hosts include rabbit (e.g., ab254718, HPA002237) , mouse (e.g., H00009957-B01P) , and goat (e.g., ABIN6742342) .
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Reactivity: Validated for human, mouse, and rat samples, with cross-reactivity in bovine and guinea pig tissues noted in some cases .
| Antibody Type | Host | Applications | Dilution |
|---|---|---|---|
| Polyclonal Rabbit | Rabbit | WB, IHC, IF | 1:500–1:2000 (WB) |
| Monoclonal Mouse | Mouse | WB, ELISA | 1:500–1:1000 (WB) |
| Polyclonal Goat | Goat | WB, IHC | 1:20–1:50 (IHC) |
Applications in Research
The HS3ST1 Antibody is employed in diverse biological studies:
2.1. Inflammation and Cardiovascular Disease
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Atherosclerosis: HS3ST1 regulates antithrombin’s anti-inflammatory activity, with a genetic variant (rs16881446) linked to increased cardiovascular risk .
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Sepsis: HS3ST1-deficient mice show exacerbated inflammation, highlighting its role in endothelial function .
2.2. Cancer Biology
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Lung Cancer: Overexpression of HS3ST1 correlates with tumor progression by inhibiting the NF-κB pathway via SPOP/FADD regulation .
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Prostate Cancer: HS3ST1 expression promotes therapeutic resistance by altering signaling pathways .
2.3. Neurodegeneration
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Alzheimer’s Disease: Increased HS3ST1 expression enhances tau pathology spread, suggesting a therapeutic target .
3.1. Atherosclerosis
3.2. Lung Cancer
| Parameter | HS3ST1 Overexpression | HS3ST1 Knockdown |
|---|---|---|
| Cell Proliferation | 2.5-fold increase in A549 cells | 40% reduction |
| Apoptosis | 60% decrease in cleaved-caspase-3 expression | 3-fold increase |
3.3. Alzheimer’s Disease
Product Specs
Target Background
- Compared to Hs3st1+/+ mice, Hs3st1-/- mice were more susceptible to LPS-induced death due to an increased sensitivity to TNF. PMID: 28126521
- Golgi-targeted HS3st1 localizes in the Golgi and results in the formation of a single type of AT-binding site and high anti-factor Xa activity. PMID: 24247246
- This study describes a conformational change that occurs in heparan sulfate 3-O-sulfotransferase-1 upon binding to heparan sulfate. PMID: 15096036
Q&A
What experimental validation methods are recommended for HS3ST1 antibody specificity?
To ensure antibody specificity in academic research, researchers should prioritize knockout controls and negative controls. For instance, in studies analyzing HS3ST1’s role in non-small-cell lung cancer (NSCLC), using Hs3st1 knockout mice or siRNA-transfected cell lines as negative controls is critical . Additionally, manufacturers like Proteintech recommend validating antibodies through antigen retrieval protocols (e.g., TE buffer pH 9.0 or citrate buffer pH 6.0 for IHC) and comparing results with positive controls such as human heart or kidney tissues .
Data Table 1: Antibody Validation Approaches
How do I optimize HS3ST1 antibody dilution for Western Blot and Immunohistochemistry?
Basic Optimization:
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Western Blot (WB): Start with a 1:500–1:1,000 dilution for human/mouse/rat samples, as validated in NSCLC cell lines (e.g., A549, H1650) .
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Immunohistochemistry (IHC): Use 1:20–1:200 dilutions, with antigen retrieval steps critical for human heart/kidney tissues .
Advanced Considerations:
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Cross-reactivity: Use blocking peptides if nonspecific binding occurs, particularly in species with conserved HS3ST1 sequences (e.g., human/mouse) .
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Signal enhancement: For low-abundance targets, amplify signals with secondary antibodies conjugated to HRP or fluorescent dyes .
Data Table 2: Antibody Dilution Guidelines
| Application | Recommended Dilution | Tested Species |
|---|---|---|
| Western Blot | 1:500–1:1,000 | Human, Mouse, Rat |
| Immunohistochemistry | 1:20–1:200 | Human Heart/Kidney |
| Immunofluorescence | 1:100–1:500 | NSCLC Cell Lines |
What are the key considerations when using HS3ST1 antibodies for co-immunoprecipitation (Co-IP) studies?
To study HS3ST1 interactions (e.g., with Glypican 4 [GPC4] in lung adenocarcinoma ), ensure:
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Antibody avidity: Use polyclonal antibodies (e.g., Proteintech 14358-1-AP) for better capture efficiency .
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Experimental controls: Include mock IP (no antibody) and input lysates to confirm pull-down specificity .
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Cell line selection: Choose models with high endogenous HS3ST1 (e.g., A549, H1650) to minimize background .
Troubleshooting Tip: If Co-IP yields weak signals, optimize lysis buffer composition (e.g., reduce SDS concentration to preserve protein-protein interactions) .
How do I resolve conflicting data on HS3ST1’s role in inflammation vs. cancer?
Conflicting outcomes often arise from context-dependent functions. For example:
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Inflammation: Hs3st1−/− mice show exacerbated septic shock due to HS3ST1’s role in antithrombin-mediated anti-inflammatory signaling .
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Cancer: HS3ST1 promotes NSCLC progression via SPOP/FADD/NF-κB pathway regulation .
Resolution Strategies:
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Experimental models: Compare findings across in vitro (cell lines), in vivo (mouse models), and human tissue studies .
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Biomarker analysis: Quantify HS3ST1 mRNA/protein levels in disease-specific contexts (e.g., NSCLC vs. AD brains) .
Data Table 3: Context-Dependent HS3ST1 Functions
| Disease | Pathway/Interaction | Experimental Model |
|---|---|---|
| NSCLC | SPOP/FADD/NF-κB | A549, H1650 cells |
| Alzheimer’s Disease | Tau internalization | AD brain tissue |
| Sepsis | Antithrombin anti-inflammatory | Hs3st1−/− mice |
What advanced techniques can I use to study HS3ST1’s role in heparan sulfate (HS) biosynthesis?
To link HS3ST1 expression to HS structural changes, employ:
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LC-MS/MS: Quantify 3-O-sulfated HS domains (e.g., Tetra-1 motif) in AD brains, as HS3ST1 knockout mice lack this domain .
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Chemoenzymatic synthesis: Generate 13C-labeled HS calibrants to detect low-abundance 3-O-sulfated HS in complex biological samples .
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CRISPR editing: Generate HS3ST1-knockout cell lines to study HS-dependent signaling pathways (e.g., Wnt, FGF) .
Data Table 4: HS3ST1-Dependent HS Analysis Methods
How do I interpret HS3ST1 antibody data in the context of Alzheimer’s disease (AD) pathology?
HS3ST1 overexpression in AD correlates with increased 3-O-sulfated HS, which facilitates tau protein internalization . Critical steps include:
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Sample selection: Compare post-mortem AD brains with pre-AD and other tauopathies .
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Structural analysis: Use LC-MS/MS to quantify HS domains (e.g., Tetra-1) that bind tau .
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Functional assays: Test synthetic 3-O-sulfated HS oligosaccharides (e.g., 14-mers) for tau uptake inhibition .
Contradiction Note: HS3ST1’s role in AD may differ from its anti-inflammatory effects in sepsis, emphasizing the need for disease-specific validation .
What controls are essential for HS3ST1 ELISA assays?
To ensure ELISA reliability, include:
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Standard curves: Use recombinant HS3ST1 protein with known concentrations .
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Negative controls: Untransfected cell lysates or non-AD brain extracts .
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Interference tests: Confirm no cross-reactivity with HS3ST1 homologs (e.g., HS3ST2) using antibody blocking peptides .
Data Table 5: ELISA Control Requirements
How can I integrate HS3ST1 antibody data with transcriptomic/proteomic profiles?
Basic Integration:
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Correlation analysis: Use Pearson’s correlation to link HS3ST1 mRNA/protein levels with clinical outcomes (e.g., NSCLC survival) .
Advanced Integration:
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Network analysis: Map HS3ST1 interactions (e.g., GPC4, SPOP) using Co-IP/MS data and bioinformatics tools (e.g., STRING) .
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Multi-omics: Combine HS3ST1 LC-MS/MS data with RNA-seq to identify HS-dependent signaling pathways in AD .
