Nanog Rabbit mAb - Q80Z64

Core Product Specifications and Parameters

Biological Background: Homeobox protein NANOG Function and Localization

• Homeobox protein NANOG (also known as ES cell-associated protein 4, Early embryo specific expression NK-type homeobox protein, Homeobox transcription factor Nanog) is a transcription regulator crucial for inner cell mass and embryonic stem (ES) cell proliferation and self-renewal (PubMed:25825768). Related references: PMID:25825768
• Imposes pluripotency on ES cells and prevents their differentiation towards extraembryonic endoderm and trophectoderm lineages. Blocks bone morphogenetic protein-induced mesoderm differentiation of ES cells by physically interacting with SMAD1 and interfering with coactivator recruitment to SMAD transcriptional complexes.
• Acts as both a transcriptional activator and repressor. Binds optimally to the DNA consensus sequences 5'-TAAT[GT][GT]-3' or 5'-[CG][GA][CG]C[GC]ATTAN[GC]-3'. It binds to the POU5F1/OCT4 promoter.
• Autorepresses its own expression in differentiating ES cells: upon interaction with ZNF281/ZFP281, it recruits the NuRD complex to its own promoter, leading to transcriptional repression.
• When overexpressed, promotes cells to enter S phase and proliferation.
• Subcellular localization is strictly nuclear.
• Tissue specificity: not expressed in oocytes and spermatogonia (at protein level) and absent from many somatic organs, ovary, testis, fibroblast, and hematopoietic cell lines.
• Alternative splicing and key post-translational modifications (e.g., phosphorylation) may regulate its activity; classified under keywords including DNA-binding, homeobox, activator, repressor, developmental protein, and reference proteome.

Experimental Guidance and Technical Tips

• The immunogen covers amino acids 160–305 of mouse Nanog, which encompasses the homeobox domain and adjacent regions. This region is highly conserved between mouse and human, supporting the observed cross-reactivity. Verify specificity in your model system.
• For Western blot, a specific band at ~34 kDa is expected in mouse ES cell lysates. Use appropriate positive and negative controls (e.g., NIH/3T3 as negative) to confirm antibody performance.
• In ChIP assays, this antibody may be suitable for enriching Nanog-bound genomic loci such as the POU5F1 promoter; consider cross-linking optimization and antibody titration.
• For intracellular flow cytometry, ensure adequate permeabilization and use proper isotype controls. Titrate the antibody to determine optimal concentration for your cell type.

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