HSD6 Antibody
Description
Target Protein: HSP60
HSP60 is a 60 kDa mitochondrial chaperonin involved in protein folding, immune modulation, and stress adaptation . It is ubiquitously expressed across species and plays roles in:
Antibody Characteristics
HSD6 Antibody (HSP60 D6F1) is a rabbit-derived monoclonal IgG antibody validated for multiple applications :
| Property | Details |
|---|---|
| Host Species | Rabbit |
| Clonality | Monoclonal (D6F1 clone) |
| Isotype | IgG |
| Applications | Western Blot (WB), Immunohistochemistry (IHC), Immunofluorescence (IF), Flow Cytometry (F) |
| Cross-Reactivity | Human, Mouse, Rat, Hamster, Monkey, Xenopus, Zebrafish, Bovine, Pig |
| Molecular Weight | 60 kDa (endogenous detection) |
This antibody demonstrates specificity for HSP60 across diverse species, with no observed cross-reactivity to mammalian HSP60 homologs in macrophage extracts .
Pathogen Neutralization
In Histoplasma capsulatum studies, IgG1/IgG2a anti-HSP60 monoclonal antibodies (e.g., clones 11D1, 6B7, 12D3) significantly:
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Prolonged survival in infected mice (60% survival rate vs. controls) .
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Reduced intracellular fungal survival by enhancing phagolysosomal fusion in macrophages .
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Increased Th1-associated cytokines (IL-2, IL-12, TNF-α) while suppressing Th2 cytokines (IL-4, IL-10) .
Diagnostic and Therapeutic Potential
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Cancer Research: HSP60 is overexpressed in tumors, making it a biomarker target for antibody-based diagnostics .
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Autoimmune Diseases: Anti-HSP60 antibodies are implicated in conditions like rheumatoid arthritis, though their therapeutic utility remains under investigation .
Validation and Quality Control
HSD6 Antibody’s reliability is supported by:
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KO Cell Line Testing: Superior specificity confirmation compared to traditional methods (e.g., Western Blot controls) .
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Multi-Application Validation: Consistent performance in WB, IHC, and IF .
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Vendor Partnerships: Collaborative efforts with repositories like the Developmental Studies Hybridoma Bank (DSHB) ensure renewable access and quality assurance .
Limitations and Challenges
Product Specs
Constituents: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4
Target Background
Q&A
Here’s a structured collection of FAQs tailored for academic researchers working with HSD17B6 (commonly abbreviated as HSD6) antibodies, incorporating experimental design principles, data analysis challenges, and methodological guidance:
Advanced Research Questions
Analysis framework:
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Assay standardization: Compare substrate concentrations (e.g., 1–100 μM for retinol vs. androsterone) .
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Epitope mapping: Confirm antibody targets (e.g., aa 200–300 may influence catalytic domain binding) .
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Orthogonal validation: Pair antibody-based detection with LC-MS quantification of reaction products.
Functional discrepancies:
| Study | Reported Activity | Likely Confounder |
|---|---|---|
| Carmona-Rivera et al. | Strong 3α-HSD activity | Tissue-specific isoform variants |
| Byrd et al. | Retinol dehydrogenase dominance | Co-factor availability (NAD⁺ vs. NADP⁺) |
Advanced solutions:
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Immunodepletion: Pre-clear samples with antibodies against homologous enzymes (e.g., HSD17B4).
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Structural modeling: Use AlphaFold-predicted HSD6 epitopes to design competition assays.
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Multiplexed SRM/MS: Develop signature peptides unique to HSD6 (e.g., VVIGGPAGLGR).
Cross-reactivity risk assessment:
| Homolog | Sequence Identity | Mitigation Strategy |
|---|---|---|
| HSD17B4 | 42% | Differential pH elution in IHC |
| HSD17B10 | 38% | CRISPR interference in cell models |
Data Interpretation Challenges
Integrative approach:
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qPCR/Western blot correlation: Normalize to housekeeping genes/proteins (e.g., GAPDH).
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Post-translational modifications: Perform deglycosylation assays (PNGase F treatment).
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Spatial resolution: Combine IHC with RNAscope® for co-localization analysis.
Common pitfalls:
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Antibody reactivity with truncated isoforms (e.g., Δexon 5 variants).
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False negatives due to epitope masking by protein complexes.
