Anti-PPP1R15A Rabbit Polyclonal Antibody (GADD34) for WB, IF/ICC, ELISA - O75807

Core Product Specifications and Parameters

Biological Background: PPP1R15A Function and Localization

• Protein identity: Also known as GADD34, growth arrest and DNA damage-inducible protein, and MyD116 homolog. Encoded by PPP1R15A gene (GeneID: 23645, SWISS: O75807). UniProt molecular weight ~73.5 kDa; observed ~73 kDa.
• Core function: Recruits protein phosphatase 1 (PP1) to dephosphorylate translation initiation factor eIF-2A, reversing stress-induced protein synthesis shut-off and facilitating cellular recovery from stress. Related references: PMID:26095357 PMID:26742780
• TGF-β pathway modulation: Down-regulates TGF-beta signaling by promoting PP1-mediated dephosphorylation of TGFB1. Related references: PMID:14718519
• Apoptosis: May promote apoptosis by inducing p53 phosphorylation on Ser-15. Related references: PMID:14635196
• Autophagy: Essential for sustained autophagy by tuning translation during starvation, thereby enabling lysosomal biogenesis and autophagic flux. Related references: PMID:32978159
• Viral restriction factor: Attenuates HIV-1 replication by inhibiting TAR RNA-mediated translation. Related references: PMID:31778897
• Microbial infection: Promotes enterovirus 71 replication by mediating the internal ribosome entry site (IRES) activity of viral 5'-UTR.
• Subcellular localization: Associates with the endoplasmic reticulum membrane and the mitochondrial outer membrane.
• Post-translational modifications: Subject to phosphorylation and ubiquitin-like conjugation (ubiquitination).
• Alternative splicing generates multiple transcript isoforms.

Experimental Guidance and Technical Tips

• The immunogen corresponds to amino acids 375-674 of human PPP1R15A, covering a C-terminal region. The resulting polyclonal antibody may recognize multiple isoforms or modified forms.
• For Western blot (WB), a band around 73 kDa is expected; additional bands may appear due to post-translational modifications or alternative splicing. Pre-treatment of samples with phosphatase inhibitors may help preserve phosphorylation-dependent signals.
• In IF/ICC, consider that GADD34 is localized to the ER and mitochondria; co-staining with organelle markers can aid verification. Permeabilization with detergents (e.g., 0.1% Triton X-100) is typically required.
• The antibody cross-reacts with human, mouse, and rat samples. Validate in your specific model system, and include appropriate positive and negative controls (e.g., stress-induced vs. untreated cells).
• ELISA applications may require optimization of coating antigen and detection conditions; the recombinant immunogen can serve as a positive control.

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