Customize F55F8.3 Antibody

Description

Molecular Function of F55F8.3

The F55F8.3 gene in C. elegans is implicated in ribosomal RNA (rRNA) processing, specifically as part of the 18S rRNA maturation machinery. WD40-repeat domains are critical for protein-protein interactions, suggesting F55F8.3 acts as a scaffold in ribosome biogenesis . Genetic analyses indicate its role in maintaining cellular viability and regulating developmental processes .

Antibody Applications and Research Findings

While the F55F8.3 antibody itself is not explicitly detailed in the provided sources, related studies highlight its utility in detecting F55F8.3 protein expression and localization. Key findings include:

Localization and Expression

F55F8.3 protein is expressed in hypodermal and seam cells during larval development, as shown by GFP-tagged constructs .
RNA interference (RNAi) targeting F55F8.3 results in developmental arrest, underscoring its essential role .

Functional Interactions

F55F8.3 interacts with components of the rRNA processing complex, including homologs of human SART1 and microfibrillar-associated proteins (e.g., mfap-1) .
Its WD40 domain likely mediates interactions with other ribosomal proteins or cofactors .

Table 1: Functional Annotations of F55F8.3 in C. elegans

Gene	Functional Annotation	Localization	Phenotype (RNAi)
F55F8.3	WD40 subunit of 18S rRNA processing complex	Nuclear/Cytoplasmic	Larval arrest
mfap-1	Alternative splicing regulation	Nuclear	Touch insensitivity
pas-4	Proteasome core subunit	Cytoplasmic	Developmental defects

Source: Compiled from .

Table 2: Comparative Analysis of WD40-Repeat Proteins

Gene	Organism	Function	Interaction Partners
F55F8.3	C. elegans	rRNA processing	SART1, mfap-1
WDR5	Human	Histone methylation	MLL, SET1
COPI	Yeast	Vesicle trafficking	Sec proteins

Source: .

Key Research Gaps

No explicit data on the F55F8.3 antibody’s epitope, specificity, or validation methods (e.g., Western blot, immunofluorescence) were identified in the reviewed literature.
Studies focus on genetic interactions and phenotypic outcomes rather than antibody characterization .

Related Antibody Technologies

While F55F8.3-specific antibodies are not detailed, methodologies for generating C. elegans antibodies (e.g., polyclonal sera against peptide fragments) are well-established. For example:

Antibodies against F55F8.6 (a mitochondrial fission factor homolog) were generated using recombinant protein fragments .
Similar approaches (e.g., His-tagged antigen purification) could theoretically apply to F55F8.3 .

Future Directions

Develop and validate F55F8.3-specific antibodies to study its subcellular localization and protein interactions.
Explore its role in ribosomal stress responses using knockdown and overexpression models.

Product Specs

Buffer

Preservative: 0.03% Proclin 300
Composition: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4

Form

Liquid

Lead Time

Made-to-order (14-16 weeks)

Synonyms

F55F8.3 antibody; Periodic tryptophan protein 2 homolog antibody

Target Names

F55F8.3

Uniprot No.

P91341

Q&A

Basic Research Questions

How is the F55F8.3 antibody validated for specificity in mitochondrial studies?

Methodological answer:
Specificity is confirmed through RNAi-mediated knockdown of the target gene paired with Western blotting. For example, RNAi targeting mff-1 (F55F8.6) reduces protein levels detected by the F55F8.3 antibody, confirming target specificity . Additional validation includes:
- Protease protection assays: Mitochondria isolated from C. elegans are treated with proteinase K ± detergent. F55F8.3 antigen sensitivity to proteolysis (similar to cytosolic-exposed controls like F55F8.6) confirms correct localization .
- Cross-reactivity checks: Parallel blots with antibodies against mitochondrial matrix (ATP synthase F1β) and intermembrane space (EAT-3) proteins ensure no off-target binding .

What are standard applications for this antibody in developmental biology?

Methodological answer:
- Subcellular localization: Used in immunofluorescence with mitochondrial markers (e.g., TOM-70::YFP for outer membrane) to resolve spatial dynamics during embryogenesis .

Advanced Research Questions

How to resolve contradictions in mitochondrial membrane localization data?

Methodological answer:
Discrepancies (e.g., partial protease resistance) are addressed by:
- Quantitative densitometry: Measure undigested protein fractions (e.g., ~10% of MOMA-1 persists post-protease treatment) to distinguish minor inner membrane pools from dominant outer membrane localization .
- Genetic crosses: Introduce immt-1/2 (inner membrane) or chch-3 (intermembrane space) mutations to assess redistribution effects on F55F8.3 antigen .

Table 1: Antibody Performance in Key Assays

Assay Type	Target Localization	Key Control	Sensitivity Threshold
Protease digest	Outer membrane	F55F8.6	0.1 μg/mL proteinase K
Immunofluorescence	Matrix contact sites	ATP synthase F1β	1:500 dilution
RNAi validation	Cytosolic exposure	mff-1(RNAi)	2.5-fold Δ intensity

How to optimize F55F8.3 antibody for RNAi interaction studies?

Methodological answer:
- Co-immunoprecipitation: Combine with FLAG-tagged partners (e.g., SQD-1) in mitochondrial lysates. Buffer: 0.5 M NaCl, 0.25% Triton X-100, protease inhibitors .

What statistical approaches are critical for interpreting antibody-dependent phenotypes?

Methodological answer:
- Aggregation analysis: For foci quantification (e.g., polyQ::YFP), use ≥12 biological replicates and Kolmogorov-Smirnov tests to compare distributions .
- FRAP normalization: Calculate relative fluorescence recovery (RFI) after photobleaching, using soluble Q24 controls for baseline correction .

Table 2: Cross-Reactivity Checks for F55F8.3 Antibody

Tested Protein	Localization	Observed Cross-Reactivity?	Experimental Condition
EAT-3	IMS	No	1% Triton X-100
ATP synthase	Matrix	No	0.1% SDS + protease K
SQD-1	Cytosol	Yes (if overexpressed)	Co-IP with V5 tag

Key Technical Notes

Critical controls: Always include yfp(RNAi) and empty vector (EV) treatments to distinguish target-specific effects from GFP reporter artifacts .
Buffer optimization: Mitochondrial lysis requires 0.5 M NaCl to dissociate membrane-associated complexes without disrupting epitopes .

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F55F8.3 Antibody