ABCI10 Antibody

Definition and Overview of ABCI10 Antibody

The ABCI10 antibody is a polyclonal antiserum developed to detect and study the ATP-binding cassette (ABC) transporter ABCI10 protein in Arabidopsis thaliana. ABC transporters are integral membrane proteins involved in ATP-dependent transport of substrates across cellular membranes. ABCI10 localizes to chloroplast inner envelope (IE) membranes and is implicated in plant-specific metabolic processes .

Key Features:

• Molecular Weight:

  • Recombinant mature At-ABCI10 runs at 27 kDa on SDS-PAGE .

  • Endogenous ABCI10 in Arabidopsis chloroplast IE membranes aligns with this size .

• Subcellular Localization:

  • Attaches to chloroplast IE membranes despite lacking transmembrane domains .

  • Resists solubilization by high ionic strength (1M NaCl) or alkaline (pH 11.3) treatments, indicating strong membrane association .

Comparative Analysis with ABCI11:

Immunoblot and Localization Studies:

• Antibody Generation:

  • Antisera were raised in rabbits against recombinant mature At-ABCI10 purified from E. coli .

  • Specificity confirmed using Arabidopsis abci10 knockout mutants .

• Membrane Fractionation:

  • ABCI10 remained membrane-bound after treatments with urea, Triton X-100, or sodium carbonate (pH 11.3) .

  • Contrasts with loosely associated proteins like Tic62, which detach under high pH .

Functional Insights:

• Role in Chloroplasts:

  • ABCI10 and ABCI11 form a prokaryote-type ECF/ABC transporter module, potentially involved in substrate allocation across IE membranes .

  • Unlike typical soluble ATPases, ABCI10’s membrane attachment suggests a regulatory or structural role .

Applications and Limitations

• Research Use:

  • Critical for studying chloroplast membrane dynamics and ABC transporter functions in plants.

  • Validated for Western blot (WB) and immunoprecipitation (IP) in Arabidopsis and pea (Pisum sativum) samples .

• Limitations:

  • Cross-reactivity with other chloroplast proteins (e.g., pea IE fractions show minor unspecific bands) .

  • Dependency on recombinant protein expression for antiserum production .

References

1. Frontiers in Plant Science (2019): Characterization of ABCI10/11 in chloroplast IE membranes .

2. Academic research on antibody validation and functional assays .