AIR9 Antibody

Introduction to AIR9 Antibody

The AIR9 antibody targets the AIR9 protein, a microtubule-associated protein (MAP) first identified in Arabidopsis thaliana. AIR9 is involved in cytokinesis, cortical division site (CDS) establishment, and cross-wall maturation . The antibody enables researchers to investigate AIR9's interactions, localization, and regulatory mechanisms through techniques like immunoprecipitation (IP), yeast two-hybrid (Y2H) screens, and fluorescence microscopy .

Key Applications of AIR9 Antibody

• Immunoprecipitation: Used to confirm physical interactions between AIR9 and other proteins, such as the kinesin KCBP .

• Localization Studies: Visualizes AIR9's dynamic distribution during the cell cycle, including its association with the preprophase band (PPB) and phragmoplast .

• Functional Analysis: Determines AIR9's role in cytokinesis and nuclear positioning in organisms like Trypanosoma brucei .

Interaction with KCBP in Arabidopsis

AIR9 interacts with the kinesin-like calmodulin-binding protein (KCBP) via its repeated A9 domains. Key experiments include:

Role in Cytokinesis and Division Site Memory

• AIR9 localizes to the PPB during G2 phase and reappears at the cortical division site during phragmoplast expansion .

• In Trypanosoma brucei, AIR9 homologs are essential for accurate cleavage furrow placement and nuclear positioning .

Immunoprecipitation Protocol

1. Extract Preparation: Homogenize Arabidopsis suspension cells in IP buffer.

2. Antibody Incubation: Mix extracts with rabbit anti-AIR9 antibody for 1 hour.

3. Protein A Beads: Add beads, wash, and elute proteins for SDS-PAGE.

4. Detection: Probe blots with anti-KCBP antibodies.

Localization in T. brucei

• AIR9-like protein (TbAIR9) localizes to cytoskeletal structures.

• RNAi knockdown disrupts cytokinesis, highlighting its functional conservation.