F09G2.8 Antibody

Here’s a structured FAQ collection for researchers working with the F09G2.8 Antibody, optimized for academic research rigor and experimental design considerations:

What are the validated applications for F09G2.8 Antibody in C. elegans studies?

The antibody is confirmed for ELISA and Western blot (WB) applications, specifically for detecting the F09G2.8 protein in Caenorhabditis elegans lysates . For WB:

• Use 10–20 µg of total protein per lane with a recommended dilution of 1:500–1:2000.

• Validate via knockdown/knockout controls (e.g., RNAi-treated worms) to confirm band specificity .

How should F09G2.8 Antibody be stored to ensure stability?

• Short-term: Store at –20°C in 50% glycerol buffer.

• Long-term: Aliquot and store at –80°C to avoid freeze-thaw cycles, which degrade IgG integrity .

• Critical: Avoid preservative-free buffers, as 0.03% Proclin 300 is required to prevent microbial growth .

What controls are essential for specificity validation?

• Positive control: Recombinant F09G2.8 protein (Uniprot O17405) .

• Negative controls:

  • Lysates from C. elegans mutants with F09G2.8 deletion .

  • Pre-immune serum from the same rabbit host to rule out nonspecific binding .

How to resolve inconsistent WB signals across developmental stages of C. elegans?

• Hypothesis: Temporal expression variability or post-translational modifications.

• Method:

  • Perform RNAi knockdown of F09G2.8 and compare protein levels via qPCR .

  • Treat lysates with phosphatase/ubiquitinase inhibitors during extraction to stabilize epitopes .

  • Use 2D gel electrophoresis to detect isoform shifts .

Can F09G2.8 Antibody cross-react with homologous proteins in other nematodes?

• Risk: High due to conserved domains in C. briggsae homologs .

• Mitigation:

  • Pre-adsorb the antibody with lysates from non-target species.

  • Perform phylogenetic alignment of F09G2.8 homologs to identify divergent regions .

How to optimize immunoprecipitation (IP) for low-abundance targets?

• Protocol adjustments:

  • Use crosslinkers like DSS to stabilize antibody-antigen interactions.

  • Combine with SILAC labeling to enhance detection sensitivity in mass spectrometry .

Data Contradiction Analysis

Integration with Omics Workflows

• Transcriptomic correlation: Pair WB data with RNA-seq from modENCODE to assess F09G2.8 expression dynamics .

• Proteomic mapping: Combine IP-MS with BioID to identify interacting partners in C. elegans lipid rafts .

Key Technical Notes

• Antibody dilution must be titrated empirically due to lot-to-lot variability .

• For long-term storage, avoid PBS-containing buffers; glycerol alone minimizes hydrolysis .