Anti-PHF8 Monoclonal Antibody for WB, ELISA - Q9UPP1

Core Product Specifications and Parameters

Biological Background: PHF8 Function and Localization

• PHF8 (KDM7B/JHDM1F/ZNF422) is a histone lysine demethylase that preferentially removes mono- and dimethyl marks from H3K9 and H4K20, converting repressive chromatin to an active state. Related references: PMID:20101266, PMID:20208542, PMID:20346720
• It is crucial for cell cycle progression by controlling the G1-S transition, and for ribosomal DNA transcription as a coactivator of RNA polymerase I. Related references: PMID:20622854, PMID:20531378
• The demethylase activity towards H3K9me2 is strongly stimulated by binding to trimethylated H3K4 (H3K4me3), providing a readout of active chromatin. Related references: PMID:20023638, PMID:20346720, PMID:20421419
• PHF8 synergizes with transcription factor ARX to upregulate KDM5C, promoting H3K4me3 enrichment at regulatory elements. Related references: PMID:31691806
• The protein is localized to the nucleus and nucleolus, in line with its chromatin and rDNA regulatory roles.
• It possesses a PHD finger that binds H3K4me3, a JmjC catalytic domain, and a zinc-finger domain (ZNF422), classifying it as a chromatin regulator.
• Required for normal brain development; loss-of-function mutations are linked to X-linked intellectual disability (MRXSSD).

Experimental Guidance and Technical Tips

• The immunogen used is a synthetic peptide corresponding to amino acids 800-900 of human PHF8, suggesting that the antibody may recognize the C-terminal region of the protein.
• For Western blot, PHF8 has a predicted molecular weight of approximately 118 kDa; ensure proper gel percentage and transfer conditions for high-molecular-weight proteins.
• Given its nuclear localization, nuclear extraction or whole-cell lysates are recommended for detection; consider validating in human and mouse cell lines that express PHF8 (e.g., HeLa, HEK293, mouse embryonic fibroblasts).
• For ELISA, the direct or sandwich format may be used; optimization of coating antigen and antibody concentration is advised.
• As a histone modifier with cell-cycle-dependent expression, signal intensity may vary; include appropriate loading controls and normalize to cell number or total protein.

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