KO Validated Anti-ATG4D Rabbit Polyclonal Antibody for WB and ELISA - Q86TL0

Core Product Specifications and Parameters

Biological Background: ATG4D Function and Localization

• ATG4D (also known as APG4D, AUTL4, autophagin-4, or HsAPG4D) is a member of the ATG4 cysteine protease family that plays a central role in autophagy.
• Proteolytic activation: ATG4D cleaves the C-terminal amino acid of ATG8 family proteins, including MAP1LC3 and GABARAPL2, to expose a glycine essential for phosphatidylethanolamine (PE) conjugation and subsequent membrane insertion (PubMed:21177865). Related references: PMID:21177865
• Delipidation activity: In addition to activation, ATG4D catalyzes the delipidation of PE-conjugated ATG8 proteins during macroautophagy and removes phosphatidylserine (PS) conjugates in non-canonical autophagy (PubMed:29458288, PubMed:33909989). Related references: PMID:29458288 PMID:33909989
• Delipidation specialization: Compared to ATG4A, ATG4B, and ATG4C, ATG4D is the major delipidation enzyme for ATG8 family proteins, with comparatively weak proteolytic activation activity.
• Mitophagy role: ATG4D promotes phagophore expansion during mitophagy by regulating ATG9A trafficking to mitochondria and facilitating endoplasmic reticulum–phagophore contacts, independent of its protease function (PubMed:33773106). Related references: PMID:33773106
• Apoptosis linkage: The mitochondrial import of ATG4D under cellular stress or differentiation links mitochondrial dysfunction with apoptosis, influencing mitochondrial physiology, ROS, and cell viability.
• Subcellular localization: ATG4D resides in the cytoplasm and can translocate to the mitochondrial matrix.
• Tissue expression: It is widely expressed in testis, and associated with processes such as apoptosis, autophagy, and thiol protease-mediated protein transport.

Experimental Guidance and Technical Tips

• The immunogen (amino acids 1–230 of human ATG4D) covers the N-terminal catalytic domain, which is highly conserved across human, mouse, and rat, consistent with the reported cross-reactivity. Consider using species-specific control lysates to confirm reactivity in your assay.
• For Western blot (WB) applications, the expected molecular weight is approximately 53 kDa. Since ATG4D expression may vary depending on cell type and stress conditions, we recommend validating the antibody in your specific sample matrix.
• The antibody has been knockout (KO) validated in certain cell lines, providing strong evidence of specificity. Nevertheless, performing your own validation with appropriate positive and negative controls is advisable.
• In ELISA, using the recombinant immunogen as a coating antigen or standard can help optimize assay conditions. Titrate the antibody to determine the optimal working concentration for your plate format.

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