At1g47790 Antibody

FAQs for Researchers: AT1 Receptor Antibodies in Academic Research
Compiled from peer-reviewed studies on antibody validation, engineering, and clinical applications

Basic: What controls are critical for Western blot analysis of AT1 receptors?

• Required Controls:

  • Include tissues/cells with confirmed AT1 receptor expression (e.g., transfected HEK293 cells) and KO models .

  • Validate against recombinant AT1 receptor protein (predicted 43 kDa band) .

  • Use secondary antibody-only lanes to rule out nonspecific binding .

Advanced: How to resolve contradictory results in AT1 receptor localization studies?

• Conflict Resolution Strategy:

  • Cross-validate using orthogonal methods:

    • Flow cytometry for membrane localization

    • Radioligand displacement assays for functional confirmation

  • Analyze antibody lot variability and storage conditions (e.g., freeze-thaw cycles) .

  • Employ epitope mapping to confirm target recognition .

Advanced: What engineering strategies improve therapeutic AT1 antibody stability?

• IgG4-Specific Modifications:

  • Introduce S228P and R409K mutations to prevent half-antibody exchange .

  • Use knob-into-hole technology for bispecific antibody design .

  • Add L235E mutation to reduce FcγR binding and off-target effects .

Key Recommendations for Researchers:

• Prioritize functional assays (e.g., ligand blocking , radioligand binding ) over immunoreactivity alone.

• Use IgG1 frameworks for effector function-dependent studies .

• Report antibody clonality, dilution, and validation data using MIABIS standards.