KRT38 Antibody
Description
Overview of KRT38 Antibody
KRT38 antibodies are immunodetection reagents targeting keratin 38, a 456-amino-acid protein (50.5 kDa) encoded by the KRT38 gene located on chromosome 17q21.2. These antibodies are essential for studying epithelial biology, particularly in dermatology and regenerative medicine .
Key Features of KRT38 Antibodies
Research Applications
KRT38 antibodies enable critical insights into epithelial cell dynamics:
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Western Blot: Detects KRT38 at ~51 kDa in human cell lines (e.g., MCF-7, HeLa, HepG2) .
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ELISA: Quantifies protein levels in tissue lysates, with dilutions ranging from 1:2,000 to 1:10,000 .
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Immunohistochemistry: Localizes KRT38 in hair follicles and nail beds, aiding studies of keratinopathies .
Recent Findings
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Mechanical Stress Response: KRT38 phosphorylation (Ser43) regulates autophagy in intervertebral disc cells under compression, linking its dysfunction to degenerative disc disease .
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Epithelial Differentiation: Co-expression with KRT5/KRT14 and KRT1/KRT10 pairs highlights its role in stratified epithelia .
Gene and Protein Details
| Attribute | Value |
|---|---|
| UniProt ID | O76015 |
| NCBI Gene ID | 8687 |
| Protein Family | Intermediate filament |
| Key Domains | α-helical rod, head/tail domains |
KRT38 heterodimerizes with type II keratins (e.g., KRT82) to form hair shaft filaments. Its dysregulation is implicated in hair disorders and epithelial fragility .
Validation and Citations
Top validated KRT38 antibodies include:
Emerging Research Directions
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Therapeutic Targets: Overexpression of KRT38 in nucleus pulposus cells reduces apoptosis under mechanical load, suggesting therapeutic potential for disc degeneration .
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Single-Cell Analysis: Single-cell RNA sequencing reveals KRT38 co-expression with KRT14 in basal epidermal cells, highlighting its role in epithelial stratification .
Limitations and Considerations
Product Specs
Composition: 50% Glycerol, 0.01M PBS, pH 7.4
Q&A
What experimental applications are validated for KRT38 antibodies in human tissue studies?
KRT38 antibodies are primarily validated for Western blot (WB) and enzyme-linked immunosorbent assay (ELISA) in human samples . For WB, recommended dilutions range from 1:1000 to 1:5000, while ELISA protocols typically use dilutions between 1:2000 and 1:10,000 . Specific validation data from independent studies show a predicted band size of 51 kDa for KRT38 in lysates from MCF-7, HeLa, and HepG2 cell lines . Researchers should confirm antibody specificity using:
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Knockout controls (e.g., CRISPR-edited KRT38-deficient cell lines)
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Peptide blocking experiments with the immunogen (recombinant human KRT38 protein, 2-456AA)
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Cross-reactivity assessments against other type I keratins (e.g., KRT37) .
How does KRT38’s molecular function inform its role in epithelial differentiation studies?
KRT38, a type I acidic keratin, heterodimerizes with type II keratins to form intermediate filaments critical for epithelial cell integrity . Key functional attributes include:
Methodologically, researchers studying epithelial differentiation should:
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Co-stain with type II keratins to confirm filament assembly
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Use calcium-switch assays to evaluate barrier function in KRT38-depleted cells .
What strategies resolve discrepancies in KRT38 expression data across publications?
Contradictory expression findings often stem from:
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Tissue-specific isoform variation: KRT38 shows differential splicing in hair follicles vs. nail beds .
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Antibody validation gaps: Only 12% of commercial KRT38 antibodies are validated via knockout controls .
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Fixation artifacts: Prolonged formalin exposure masks epitopes; antigen retrieval with pH 9.0 Tris-EDTA improves detection .
To harmonize data:
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Standardize lysis buffers (RIPA vs. NP-40 impacts keratin solubility)
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Validate across multiple platforms (e.g., WB, immunohistochemistry, RNA-seq)
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Use reference cell lines (e.g., HaCaT for epidermal studies) .
How can KRT38 antibodies be optimized for co-immunoprecipitation (Co-IP) studies?
Successful Co-IP requires:
Notably, KRT38 forms obligate heteropolymers; Co-IP with type II keratins (e.g., KRT85) confirms functional interactions .
What phylogenetic insights guide cross-species KRT38 studies?
KRT38 exhibits lineage-specific conservation:
| Species | Identity (%) | Functional equivalence |
|---|---|---|
| Human | 100 | Native |
| Chimpanzee | 98.7 | Preserved |
| Bovine | 82.1 | Partial |
| Zebrafish | 47.6 | Divergent |
For translational models:
How do post-translational modifications impact KRT38 antibody binding?
Common modifications affecting epitope recognition:
| Modification | Site | Effect on Antibody Binding |
|---|---|---|
| Phosphorylation | Ser-32 | Reduces affinity by 60% |
| O-GlcNAcylation | Thr-214 | Epitope masking |
| Proteolysis | N-terminal | False-negative WB results |
Mitigation strategies:
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Treat lysates with phosphatase inhibitors (e.g., PhosSTOP)
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Use deglycosylation enzymes (e.g., PNGase F) during sample prep .
What multiplex approaches enhance KRT38 pathway analysis?
A validated 8-plex panel for epithelial studies:
| Target | Assay | Purpose |
|---|---|---|
| KRT38 | Luminex | Core filament quantification |
| KRT85 | MSD-ECL | Heterodimer partner |
| Caspase-14 | ELISA | Differentiation marker |
| TGM3 | WB | Cross-linking enzyme |
Normalize data to housekeeping proteins (e.g., β-actin) and account for batch effects via ComBat .
How should researchers validate KRT38 antibodies for single-cell RNA-seq correlation?
Follow this workflow:
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scRNA-seq: Cluster epithelial cells using KRT38 expression (cutoff: >5 TPM) .
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Immunofluorescence: Compare protein localization to transcript hotspots.
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Spatial transcriptomics: Resolve discrepancies using 10x Visium (≥5,000 spots/mm²) .
Key quality metrics:
How do KRT38 mutations influence antibody performance in genodermatosis research?
Pathogenic variants (e.g., p.Arg104Cys) cause:
Solution pipeline:
