FLA8 Antibody

Definition and Target

FLA8 antibody is a rabbit-derived polyclonal antibody that specifically recognizes the FLA8 protein subunit of heterotrimeric kinesin-II, a motor protein complex critical for cargo transport along cilia and flagella. This antibody is primarily utilized to investigate IFT mechanisms in model organisms such as Chlamydomonas reinhardtii .

Key Research Applications

FLA8 antibody has been employed in multiple experimental techniques to elucidate kinesin-II functionality:

Source:

Role in Intraflagellar Transport

Heterotrimeric kinesin-II, composed of FLA8 (KIF3B homolog), FLA10 (KIF3A homolog), and a accessory subunit, drives anterograde transport of IFT particles from the ciliary base to the tip. Disruption of FLA8 impairs ciliary assembly and function, as shown in Chlamydomonas mutants .

Ciliary Dynamics

Studies using FLA8 antibody have revealed that kinesin-II is essential for:

• Maintenance of ciliary length and stability.

• Transport of tubulin and other structural proteins to growing ciliary tips.

• Coordination with dynein motors for bidirectional IFT regulation.

Reactivity

FLA8 antibody exhibits specificity for conserved epitopes in:

• Chlamydomonas reinhardtii (primary model organism).

• Other eukaryotic systems with ciliary/kinesin-II homologs (inferred from functional conservation) .

Validation

• Positive Controls: Wild-type Chlamydomonas ciliary extracts show strong WB bands at ~85 kDa (predicted molecular weight of FLA8).

• Negative Controls: fla8 knockout mutants exhibit no detectable signal .

Functional Insights

• Temperature Sensitivity: fla8 mutants display shortened cilia at restrictive temperatures, reversible upon FLA8 reintroduction.

• IFT Particle Association: Co-localization experiments using FLA8 antibody confirm kinesin-II’s direct interaction with IFT-B complexes.

Pharmacological Interactions

• Microtubule-depolymerizing agents (e.g., nocodazole) abolish FLA8-dependent transport, confirming microtubule track reliance.

Limitations and Future Directions

While FLA8 antibody has been instrumental in foundational IFT studies, its utility in mammalian systems remains underexplored. Recent advances in cryo-EM and CRISPR-based tagging (e.g., GFP-FLA8 fusions) could reduce dependence on antibody-based detection. Further validation in metazoan models is warranted to expand its applications .