RAB11FIP4 Antibody

Basic Research Questions

• What is RAB11FIP4 and what are its primary cellular functions?

  RAB11FIP4 (Rab11 family interacting protein 4) primarily functions as a regulator of endocytic trafficking by participating in membrane delivery processes. It belongs to the class II FIP family characterized by the presence of EF-hand domains rather than phospholipid-binding C2 domains . Current research demonstrates that RAB11FIP4:

  • Acts as an effector for Rab11 GTPase in regulating endosomal recycling

  • Is required for the abscission step in cytokinesis, likely by facilitating delivery of recycling endosome membranes to the cleavage furrow

  • Participates in autophagy regulation, particularly affecting both macroautophagy and chaperone-mediated autophagy (CMA) pathways

  • Plays a role in viral egress during certain infections (e.g., HCMV), independently of ARF6

  • May influence cellular homeostasis through regulation of endoplasmic reticulum (ER) stress response

• What experimental applications are RAB11FIP4 antibodies suitable for?

  Based on validated research applications, RAB11FIP4 antibodies are suitable for multiple experimental techniques:

  Application	Validated Use	Typical Dilution Range
  Western Blotting (WB)	Detection of endogenous RAB11FIP4	1:500-1:1000
  Immunohistochemistry (IHC)	Tissue expression analysis	1:100-1:1000
  Immunocytochemistry (ICC)	Cellular localization studies	As recommended by manufacturer
  Immunofluorescence (IF)	Subcellular distribution analysis	As recommended by manufacturer
  ELISA	Protein quantification	1:40000

  When designing experiments, consider optimizing antibody concentrations based on your specific tissue/cell type and experimental conditions .

• How should researchers validate RAB11FIP4 antibodies for their specific experimental systems?

  Proper validation ensures reliable research outcomes. Implement the following methodological approach:

  • Positive controls: Use cell lines known to express RAB11FIP4, such as PANC-1 which shows moderate expression (relative expression level of 15.368 ± 0.132)

  • Negative controls: Include samples with RAB11FIP4 knocked out via CRISPR/Cas9 or siRNA silencing

  • Size verification: Confirm detection of the predicted 70 kDa band in Western blots

  • Cross-reactivity assessment: Test antibody specificity across intended species (human, mouse, etc.)

  • Application-specific controls: For IHC/ICC, include peptide competition assays and isotype controls

  • Functional validation: Confirm that antibody can detect changes in RAB11FIP4 expression levels following experimental manipulations such as gene knockout or overexpression

• What are the optimal storage and handling conditions for RAB11FIP4 antibodies?

  To maintain antibody integrity and performance:

  • For short-term storage (up to 1 week): Store at 2-8°C

  • For long-term storage: Maintain at -20°C or -80°C in small aliquots to prevent repeated freeze-thaw cycles

  • Many commercial preparations are supplied in buffered solutions containing stabilizers:

    • Typically in 1X PBS buffer with 0.09% (w/v) sodium azide and 2% sucrose or

    • 1X PBS with 20% glycerol (pH 7.0) and 0.01% thimerosal as preservative

  • Allow antibody to equilibrate to room temperature before opening

  • Centrifuge briefly before use to collect solution at the bottom of the vial

  • Follow manufacturer's recommendations for reconstitution if supplied as lyophilized powder