[KO Validated] Anti-AIF Rabbit Polyclonal Antibody for WB, IF, IHC, ELISA - O95831

Core Product Specifications and Parameters

Biological Background: AIF Function and Localization

• AIF, also known as programmed cell death protein 8 (PDCD8), is encoded by the AIFM1 gene in humans. Multiple isoforms exist due to alternative splicing, including a pro-apoptotic isoform and brain-specific variants.
• AIF functions as an NADH oxidoreductase in normal mitochondrial metabolism and as a key regulator of caspase-independent apoptosis. Upon apoptotic stimulation, it is released from the mitochondrial intermembrane space into the cytosol and nucleus, where it triggers chromatin condensation and large-scale DNA fragmentation (parthanatos). Related references: PMID:17094969 PMID:20362274 PMID:23217327
• In the nucleus, AIF binds to DNA in a sequence-independent manner and induces caspase-independent chromosomal DNA fragmentation, a hallmark of parthanatos. Related references: PMID:27178839 PMID:20362274
• AIF interacts with EIF3G to inhibit protein synthesis by blocking the EIF3 translation machinery and activates caspase-7, thereby amplifying apoptosis. Related references: PMID:17094969
• AIF plays a critical role in caspase-independent, pyknotic cell death induced by hydrogen peroxide exposure. Related references: PMID:19418225
• In mitochondrial homeostasis, AIF participates in respiratory chain biogenesis by interacting with CHCHD4 and controlling its mitochondrial import. Related references: PMID:26004228
• AIF is predominantly localized in the mitochondrial intermembrane space and inner membrane under normal conditions. During apoptosis, it translocates to the cytoplasm, perinuclear region, and nucleus.
• AIF is expressed in all tested human tissues except brain (isoform 5). It is detected in muscle, skin fibroblasts, and osteoblasts. Notably, isoform 5 is frequently down-regulated in various human cancers.

Experimental Guidance and Technical Tips

• The immunogen corresponds to the C-terminal region (aa 334-613) of human AIF, which may allow recognition of multiple splice variants. For applications such as Western blot (WB), expect a band around 67 kDa. Consider using knockout (KO)-validated lysates to confirm specificity.
• For immunofluorescence (IF/ICC), note that AIF predominantly localizes to mitochondria in healthy cells, but translocates to the nucleus upon apoptosis induction. Fixation and permeabilization conditions should be optimized to preserve both mitochondrial and nuclear staining.
• In immunohistochemistry (IHC-P), antigen retrieval may be necessary to unmask epitopes; test standard citrate or EDTA-based protocols. Validate in positive control tissues (e.g., muscle, skin) expressing AIF.
• For ELISA, the polyclonal nature allows use as both capture and detection antibody in sandwich formats, but pairing with a monoclonal may reduce background.

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