VPS2.3 Antibody

Role in ESCRT-III Machinery

VPS2.3 interacts with other ESCRT-III subunits (e.g., VPS24.1) and associated proteins like AMSH3, a deubiquitinating enzyme critical for endosomal trafficking. Mutations in VPS2.3’s MIT-interacting motif (MIM1) disrupt these interactions, highlighting its structural importance .

Species-Specific Features

In Arabidopsis thaliana, VPS2.3 differs from paralogs VPS2.1 and VPS2.2 in its MIM1 sequence, which affects binding partners and functional outcomes .

Design and Applications

While no commercial VPS2.3 antibody is explicitly documented, analogous antibodies (e.g., VPS35, VPAC2) provide a template for expected properties:

• Host Species: Typically rabbit-derived polyclonal or recombinant monoclonal antibodies .

• Applications: Western blot (WB), immunoprecipitation (IP), immunofluorescence (IF) .

• Validation: Requires knockout (KO) controls to confirm specificity .

Table 1: Hypothetical Validation Parameters for VPS2.3 Antibody

Specificity Concerns

Antibodies targeting ESCRT-III subunits often face cross-reactivity risks due to structural homology. For example:

• The anti-VPAC2 antibody SP235 detects VPAC2 in humans, mice, and rats but not VPAC1 .

• Anti-VPS35 antibodies show variability in performance across applications .

Recommendations for Use

• Knockout Validation: Essential to rule off-target binding .

• Concentration Titration: Optimize to avoid nonspecific signals .

ESCRT-III Studies

VPS2.3 antibodies would enable:

• Mapping protein-protein interaction networks (e.g., with AMSH3 or VPS24.1) .

• Investigating mutations affecting membrane scission or autophagy .

Broader Impact

Poor antibody validation contributes to reproducibility crises. Third-party testing, as proposed for hVPS35 antibodies, is critical .

Future Directions

• Recombinant Antibodies: Engineered for higher specificity (e.g., GeneTex GTX635821) .

• Multispecies Cross-Reactivity: Testing in model organisms (e.g., Arabidopsis, mice) .