Customize daz-1 Antibody

Product Specs

Buffer

Preservative: 0.03% Proclin 300
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4

Form

Liquid

Lead Time

Made-to-order (14-16 weeks)

Synonyms

daz-1 antibody; F56D1.7DAZ protein 1 antibody

Target Names

daz-1

Uniprot No.

Q20870

Target Background

Function

DAZ-1 is an RNA-binding protein that plays a crucial role in oogenesis, but not in spermatogenesis. It is essential for meiotic entry and germline differentiation, specifically at the pachytene stage of meiosis I in female germ cells, regardless of the soma's sex. DAZ-1 is believed to regulate the translation of specific mRNAs by binding to their 3'-UTR.

Gene References Into Functions

DAZ-1 in C. elegans plays critical roles in female premeiotic and early meiotic germ cells, likely by regulating the translational activity of specific target genes required for the progression of oogenesis. PMID: 15572146
daz-1 plays a role upstream of the pathway for germ cell sex determination. PMID: 16641369
In a two-hybrid assay, DAZ-1, as bait, interacted with CBP-3. PMID: 16678151
All germline samples studied were found to be free from AZFc microdeletions and CNP of the DAZ genes. PMID: 17308897

Database Links

KEGG: cel:CELE_F56D1.7

STRING: 6239.F56D1.7

UniGene: Cel.6835

Protein Families

RRM DAZ family

Tissue Specificity

Germline specific. More strongly expressed during oogenesis than during spermatogenesis. During the larval stages, it is more abundant at the distal region than the proximal region of the gonad. In young adult hermaphrodites, it is expressed at a very low

Q&A

Basic Research Questions

How to validate DAZ-1 antibody specificity for spermatogenesis studies?

Methodology:
- Perform knockdown/knockout validation: Compare Western blot signals in DAZ1-deficient cell lines (e.g., DAOY cells with DAZ shRNA ) against controls.
- Use recombinant protein controls: Validate with DAZ1 (AA 21-120) GST-tagged immunogen (SSSAAASQGW...KKLC) .
- Tissue-specific staining: Confirm cytoplasmic/nuclear localization patterns in human testis sections (Figure 2h in , IHC-P at 1:10 dilution in ).

What experimental applications are optimal for DAZ-1 antibodies?

Application	Protocol Notes	Key Findings from Literature
Western Blot	Use 20-30 µg testis lysate; detect ~35 kDa band	DAZ1 overexpression increases PABPC1 co-immunoprecipitation efficiency
Immunofluorescence	Fixation: 4% PFA; permeabilization with 0.1% Triton X-100	Nuclear-to-cytoplasmic translocation observed during spermatocyte maturation
PAR-CLIP-seq	Crosslink at 365 nm; immunoprecipitate with DAZ1 antibody	Identified 2,357 mRNAs with 3'-UTR UGUU motifs bound by DAZ1

Advanced Research Questions

How to resolve contradictions in DAZ-1 localization studies across publications?

Case Analysis:
- Nuclear localization reports: Seen in fetal gonocytes (IHC data in ).
- Cytoplasmic dominance: Observed in spermatogonia (single-cell RNA-seq in ).
- Resolution strategy:
  - Stage-specific staining: Use testis sections at differing developmental stages.
  - Fractionation assays: Separate nuclear/cytoplasmic lysates (as in polysome profiling).

What mechanisms link DAZ-1 antibody reactivity to translational regulation?

Experimental Design:
- Polysome profiling: Compare polysome/monosome (P/M) ratios in DAZ1-knockdown vs. wild-type cells:
  
  Condition P/M Ratio Key Proteins Affected
  Control 2.8 ± 0.3 RPS6, RPL11 stable
  DAZ1-KD 1.2 ± 0.2 PABPC1 dissociation
- RIP-qPCR: Immunoprecipitate DAZ1-bound mRNAs (e.g., CCND1, CDK6) and quantify enrichment (4-7x vs. IgG controls) .

Condition	P/M Ratio	Key Proteins Affected
Control	2.8 ± 0.3	RPS6, RPL11 stable
DAZ1-KD	1.2 ± 0.2	PABPC1 dissociation

How to integrate DAZ-1 antibody data with single-cell transcriptomics?

Workflow:
- Validation cohort: Compare scRNA-seq clusters (e.g., c-KIT+ spermatogonia ) with DAZ1 IHC intensity.
- Functional correlation:
  - High DAZ1 expression → 62% reduction in G1-phase cells (p < 0.01) .
  - Low DAZ1 → 40% increase in S-phase arrest .

Technical Optimization

Why do DAZ-1 antibodies show variable performance in ELISA vs. Western blot?

Troubleshooting Guide:

Issue Cause Solution
Low WB signal Conformational epitope disruption Use mild detergents (CHAPS > SDS)
High ELISA background Cross-reactivity with DAZ2-4 Pre-absorb with DAZ2/3/4 peptides

Issue	Cause	Solution
Low WB signal	Conformational epitope disruption	Use mild detergents (CHAPS > SDS)
High ELISA background	Cross-reactivity with DAZ2-4	Pre-absorb with DAZ2/3/4 peptides

How to design controls for DAZ-1 functional studies in AZFc deletion models?

Recommended Controls:
- Positive: Testis samples from OA (obstructive azoospermia) patients with intact DAZ .
- Negative: AZFc-deleted patient samples (≈20% DAZ1 expression) .
- Technical: siRNA-resistant DAZ1 rescue construct (e.g., silent mutations at shRNA target sites) .

Data Interpretation

How to distinguish DAZ1-specific effects from DAZL interference?

Experimental Approaches:
- Selective knockdown: Use siRNAs targeting DAZ1-unique 3'UTR regions.
- Co-staining: Pair DAZ1 antibody with DAZL-specific probes (distinct nuclear localization in spermatocytes ).

What statistical thresholds apply to DAZ-1 interaction proteomics?

MS Data Criteria:
- Significance: ≥2 unique peptides, FDR <1% .
- Biological relevance: Prioritize partners like PABPC1 (8-fold enrichment in IP-MS) .

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daz-1 Antibody