Customize daf-36 Antibody

Description

Biological Role of DAF-36

DAF-36 (Neverland) is a cholesterol 7,8-dehydrogenase that catalyzes the conversion of cholesterol to 7-dehydrocholesterol (7dC), a rate-limiting step in steroid hormone production . This enzyme is evolutionarily conserved across nematodes (Caenorhabditis elegans), insects (Drosophila melanogaster), and vertebrates (Danio rerio, Xenopus laevis) . Key features include:

A Rieske [2Fe-2S] domain and non-heme iron-binding motif essential for catalytic activity .
Rescue of developmental defects in daf-36 mutants via 7dC supplementation, confirming its enzymatic role .

Enzymatic Activity

In vitro assays demonstrate DAF-36’s cholesterol 7,8-dehydrogenase activity:

Species	Cholesterol Conversion to 7dC (%)	Source
C. elegans	~0.9
Bombyx mori	~2.6
D. melanogaster	Not detected*

*Functional activity confirmed via ortholog expression (e.g., D. mojavensis) .

Applications of the daf-36 Antibody

The antibody is utilized to:

Localize DAF-36 expression in tissues (e.g., prothoracic glands in insects) .
Validate protein knockdown in genetic studies via Western blot or immunohistochemistry.
Inhibit enzymatic activity to study cholesterol metabolism pathways.

Comparative Analysis Across Species

DAF-36 orthologs share functional conservation despite sequence divergence:

Species	Rescue Efficacy in D. melanogaster	Key Phenotype
C. elegans	Full rescue	Normal pupation
X. laevis	Partial rescue	Delayed metamorphosis
D. rerio	Full rescue	Restored ecdysteroid synthesis

Implications for Biomedical Research

Steroid Hormone Disorders: Mutations in daf-36 homologs may underlie defects in hormone-sensitive processes (e.g., metamorphosis, molting) .
Parasite Targeting: Since daf-36 is absent in mammals, it represents a potential therapeutic target for helminth infections .

Product Specs

Buffer

Preservative: 0.03% ProClin 300
Components: 50% Glycerol, 0.01M PBS, pH 7.4

Form

Liquid

Lead Time

14-16 weeks (made-to-order)

Synonyms

daf-36 antibody; C12D8.5 antibody; Cholesterol 7-desaturase antibody; EC 1.14.19.21 antibody; Cholesterol desaturase daf-36 antibody

Target Names

daf-36

Uniprot No.

Q17938

Target Background

Function

This antibody targets DAF-36, an enzyme that catalyzes the desaturation of cholesterol to 7-dehydrocholesterol (7-DHC). This crucial step initiates the biosynthesis of Δ⁷-dafachronic acid, a key steroid hormone in nematodes. Δ⁷-dafachronic acid functions as a ligand for the nuclear hormone receptor DAF-12, regulating developmental processes such as dauer formation and reproductive growth. Inhibition of DAF-12 by Δ⁷-dafachronic acid suppresses dauer formation and promotes reproductive development.

Gene References Into Functions

DAF-36 functions as a cholesterol 7-desaturase, converting cholesterol to 7-dehydrocholesterol. PMID: 21749634

Database Links

KEGG: cel:CELE_C12D8.5

STRING: 6239.C12D8.5

UniGene: Cel.4822

Subcellular Location

Membrane; Single-pass membrane protein.

Tissue Specificity

Expressed in intestine at all postembryonic stages, including dauer. Expression is reduced in daf-2 mutants.

Q&A

Here’s a structured collection of FAQs tailored to academic research on daf-36 antibody applications, integrating experimental design principles, methodological insights, and data analysis strategies:

What experimental designs are optimal for studying daf-36’s role in transgenerational epigenetic inheritance?

Advanced Research Focus

Approach:
- Combine lifespan assays with H3K4me2 quantification across generations in spr-5 mutants .
- Use daf-36 RNAi to dissect its interaction with the DAF-12/dafachronic acid pathway .
Key Metrics:
- Track fertility decline (e.g., egg-laying capacity) and lifespan extension over 15+ generations .
- Measure daf-36 mRNA levels via qPCR to correlate epigenetic changes with phenotype .

How to resolve contradictions in daf-36’s role in H3K4me2 accumulation vs. lifespan regulation?

Data Contradiction Analysis

Observation: daf-36 knockdown eliminates lifespan extension but does not affect H3K4me2 levels in spr-5 mutants .
Hypothesis Testing:
- Use dafachronic acid supplementation to bypass daf-36 and test rescue of lifespan phenotypes .
- Perform ChIP-seq to identify H3K4me2 targets independent of daf-36.
Statistical Validation: Apply two-way ANOVA to compare treatment effects across genotypes .

What methods are recommended for quantifying daf-36-dependent steroid signaling in longevity studies?

Methodological Guidance

Assays:
- Lifespan Analysis: Compare daf-36(RNAi) vs. control worms under standardized conditions (n ≥ 100 per group) .
- Steroid Quantification: Use LC-MS to measure dafachronic acid levels in late-generation mutants .
Table: Key Experimental Findings

Parameter	spr-5 Mutants (G15)	spr-5 + daf-36(RNAi)	Wild Type + DA Supplementation
Median Lifespan Extension	30.2%	3.9%	14.5%
H3K4me2 Level	Elevated	No change	Baseline
daf-36 mRNA Expression	2.5-fold increase	0.8-fold	1.0-fold

How to design multi-omics studies linking daf-36 to epigenetic and metabolic pathways?

Advanced Integrative Research

Workflow:
- Perform RNA-seq to identify daf-36-regulated genes (e.g., DAF-12 targets) .
- Integrate ATAC-seq data to map chromatin accessibility changes.
- Validate findings with targeted metabolomics (steroid profiling) .
Troubleshooting:
- Address batch effects in transgenerational studies by synchronizing worm populations.
- Use permutation testing to account for multiple comparisons in omics datasets.

What controls are critical for daf-36 antibody-based assays in genetic screens?

Quality Control Strategies

Technical Controls:
- Include daf-36 overexpression strains to test antibody linearity.
- Validate cross-reactivity using daf-12 or daf-16 mutants .
Biological Controls:
- Compare phenotypes in daf-36(RNAi) vs. daf-12 ligand-binding domain mutants .

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daf-36 Antibody