lin-26 Antibody
Description
Introduction to LIN-26 Antibody
The LIN-26 antibody is a polyclonal reagent used to detect the LIN-26 protein in C. elegans, a zinc-finger transcription factor critical for specifying non-neuronal cell fates. LIN-26 is expressed in hypodermal cells, glial-like socket/sheath cells, and somatic gonad precursors, where it represses neuronal gene expression to maintain epithelial identity .
Molecular Characterization of the LIN-26 Antibody
The antibody targets the first two-thirds of the LIN-26 protein, excluding its zinc-finger domains. Key properties include:
| Property | Details |
|---|---|
| Target Epitope | N-terminal region (excluding zinc fingers) of LIN-26 (53.2 kDa predicted) |
| Reactivity | C. elegans nuclear antigens (hypodermal, socket/sheath, somatic gonad cells) |
| Detection Methods | Immunohistochemistry (IHC), Western blotting (WB) |
| Observed Bands (WB) | 55 kDa and 58 kDa polypeptides (possible isoforms or post-translational modifications) |
Applications in Research
The LIN-26 antibody is instrumental in developmental biology studies, enabling precise localization of LIN-26 in:
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Hypodermal Cells: Stains nuclei of seam cells, lateral hypodermis, and rectal cells .
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Sensory Organs: Identifies socket/sheath cells supporting amphid, phasmid, and cephalic neurons .
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Somatic Gonad: Labels Z1/Z4 germline precursors and uterine cells .
Key Findings from LIN-26 Antibody Studies:
Technical Protocols and Limitations
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IHC Protocol:
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Limitations:
Functional Insights from LIN-26 Antibody Research
The antibody has elucidated LIN-26’s role in:
Product Specs
**Constituents:** 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
STRING: 6239.F18A1.2.2
UniGene: Cel.38780
Q&A
Basic Research Questions
How to validate LIN-26 antibody specificity in C. elegans developmental studies?
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Perform Western blotting against lin-26 mutant lysates (e.g., lin-26(n156)) to confirm absence of signal .
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Combine immunofluorescence with genetic mutants: compare staining patterns in wild-type vs. hypodermal/glial cell-deficient strains .
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Use RNAi knockdown followed by antibody staining to observe signal reduction in target tissues.
What experimental designs leverage LIN-26 antibody to study non-neuronal cell fate specification?
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Time-course staining: Track LIN-26 protein dynamics during embryogenesis using synchronized worm populations .
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Cell lineage analysis: Combine antibody staining with fluorescent reporters (e.g., lin-26p::GFP) to correlate protein localization with cell fate decisions.
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Mutant rescue experiments: Express LIN-26 under tissue-specific promoters and validate restoration of hypodermal/glial markers via antibody co-staining.
What controls are essential for LIN-26 antibody-based assays?
Advanced Research Questions
How to resolve contradictions in LIN-26 expression patterns across studies?
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Quantitative imaging: Measure fluorescence intensity ratios between hypodermal vs. neuronal cells to assess asymmetric segregation .
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Conditional mutants: Use temperature-sensitive alleles to distinguish primary vs. secondary effects on protein stability.
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Multi-omics integration: Cross-validate antibody data with single-cell RNA-seq of lin-26-expressing cells.
What methodologies detect LIN-26 protein segregation during asymmetric cell divisions?
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Fixed-cell analysis: High-resolution confocal microscopy of early embryos stained with LIN-26 antibody .
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FRAP (Fluorescence Recovery After Photobleaching): Quantify protein turnover rates in daughter cells post-division.
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Co-staining with cell fate markers: Use antibodies against neuronal (e.g., UNC-86) and hypodermal (e.g., ELT-3) proteins to correlate LIN-26 loss with differentiation.
How to investigate LIN-26’s role in maintaining non-neuronal cell identity?
| Approach | Key Steps | Outcome Metric |
|---|---|---|
| ChIP-seq | Chromatin immunoprecipitation with LIN-26 antibody | Identify direct transcriptional targets |
| Proteomics | Immunoprecipitation + mass spectrometry | Map protein interaction networks |
| Live imaging | LIN-26::GFP translational reporter | Spatiotemporal dynamics in lin-26 mutants |
Data Contradiction Analysis Framework
Case Example: Discrepancies in glial cell transformation phenotypes
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Technical variables: Compare antibody lot numbers, fixation protocols (e.g., methanol vs. formaldehyde).
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Strain background: Validate mutations in linked loci using whole-genome sequencing.
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Phenotypic quantification:
Phenotype Wild-Type lin-26(n156) Glial cell death 0% 38% Neuronal transformation 0% 22%
