Customize C05D11.8 Antibody

Description

Molecular Identification and Target Protein

The C05D11.8 antibody recognizes the C. elegans protein encoded by the C05D11.8 locus (UniProt ID: Q11187) . Structural predictions using AlphaFold 3 indicate that C05D11.8 (FHIP-1) contains:

A C-terminal FHIP (FHF Complex Subunit) domain (residues 749–920)
A GTP–RAB-5 binding interface critical for endosomal trafficking

Property	Details
Organism	Caenorhabditis elegans
Target Gene	C05D11.8
Protein Homology	Human FHIP (FHF complex subunit)
Structural Domains	Coiled-coil regions, FHIP domain, GTP–RAB-5 interaction motif
Applications	Western blotting, immunoprecipitation, functional genomics studies

Functional Role in Intracellular Trafficking

C05D11.8 is a core component of the FHF complex, which regulates dynein-mediated transport of early endosomes. Key findings include:

Interaction with ZYG-12/Hook: Forms a dynein adaptor system to position early endosomes .
GTP–RAB-5 Binding: AlphaFold modeling reveals direct binding to GTP-loaded RAB-5, a master regulator of endosomal maturation .
Phenotypic Effects: RNAi-mediated depletion causes perinuclear clustering of early endosomes, disrupting organelle distribution .

Research Applications and Protocols

The C05D11.8 antibody has been utilized in:

Western Blotting: Detects endogenous FHIP-1 in C. elegans lysates under denaturing conditions .
Co-Immunoprecipitation: Validates interactions with RAB-5 and ZYG-12 in dynein adaptor studies .
Functional Genomics: Screens for migration defects in distal tip cells (DTCs) via RNAi knockdown .

Experimental Use	Conditions
Antibody Dilution	1:1000 (Western blot), 1:200 (IP)
Host Species	Rabbit (polyclonal)
Validation	Loss-of-function phenotypes confirmed via RNAi

Comparative Analysis with Influenza Antibodies

While C05D11.8 focuses on trafficking, structural parallels exist with influenza-neutralizing antibodies (e.g., C05, 5J8) :

Single-Loop Targeting: C05D11.8’s coiled-coil domains mimic the elongated CDR H3 loops in influenza antibodies that penetrate conserved viral epitopes .
Conserved Binding Mechanisms: Both systems rely on minimal footprints to engage dynamic interfaces (viral HA vs. RAB-5) .

Technical Considerations

Cross-Reactivity: No observed cross-reactivity with human FHIP in validation studies .
Limitations: Requires GTP-bound RAB-5 for stable complex formation in vitro .
Commercial Availability: Sold as a research-grade reagent (Cusabio Product Code: CSB-PA607541XA01CXY) .

Product Specs

Buffer

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

Form

Liquid

Lead Time

Made-to-order (14-16 weeks)

Synonyms

C05D11.8 antibody; UPF0518 protein C05D11.8 antibody

Target Names

C05D11.8

Uniprot No.

Q11187

Q&A

Here’s a structured FAQ collection for researchers investigating the C05D11.8 Antibody in C. elegans and related systems, synthesized from experimental methodologies and findings in the provided literature:

Advanced Research Questions

How to resolve contradictory results between Western blot and immunoprecipitation data?

Troubleshooting steps:
- Verify antibody cross-reactivity using knockout lysates ( ).
- Optimize IP buffer stringency (e.g., adjust sodium deoxycholate concentration) to reduce nonspecific interactions ( ).
- Quantify band intensities with tools like ImageJ and normalize to housekeeping proteins (e.g., GAPDH) ( ).

What strategies identify C05D11.8’s interaction partners in a tissue-specific manner?

Approach:
- Perform tissue-specific IP-MS (immunoprecipitation followed by mass spectrometry) using strains expressing tagged C05D11.8 in specific cell types ( ).
- Validate interactions using AlphaFold-predicted structural interfaces (e.g., FHIP-1/RAB-5 binding sites) and site-directed mutagenesis ( ).

How to address epitope masking in fixed samples for C05D11.8 localization?

Solutions:
- Test alternative fixation methods (e.g., methanol-free formaldehyde).
- Use antigen retrieval buffers (e.g., citrate buffer, pH 6.0) to expose hidden epitopes ( ).
- Validate with live-cell imaging of GFP-tagged C05D11.8 ( ).

Data Interpretation & Technical Challenges

How to analyze C05D11.8’s role in condensin complex regulation?

Experimental design:
- Compare phenotypes of smcl-1(0) (null) and dpy-28 (condensin hypomorph) mutants using brood size assays and chromosomal staining ( ).
- Quantify condensin I/II ratios via quantitative Western blotting with subunit-specific antibodies ( ).

What controls are critical for RNAi-based functional studies of C05D11.8?

Essential controls:

Control Type Purpose
Empty vector RNAi Assess off-target effects
gfp RNAi Confirm target specificity
Rescue experiments Validate phenotype reversibility

Control Type	Purpose
Empty vector RNAi	Assess off-target effects
gfp RNAi	Confirm target specificity
Rescue experiments	Validate phenotype reversibility

How to reconcile conflicting localization data between studies?

Analysis framework:
- Compare antibody clones and epitope tags used (e.g., Myc vs. Flag vs. endogenous).
- Assess fixation/permeabilization protocols across studies ( ).
- Validate using orthogonal methods (e.g., CRISPR-tagged strains vs. antibody staining) ( ).

Method Optimization Tables

Table 1: Antibody Storage Conditions

Condition	Stability (Months)	Source
-80°C in 50% glycerol	24
4°C in sodium azide	6

Table 2: Cross-reactivity Risk Mitigation

Strategy	Efficacy
Pre-adsorption with KO lysates	High (90% reduction)
Competitive peptide blocking	Moderate (70%)

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C05D11.8 Antibody