ZNF367 Antibody

What is ZNF367 and what are its primary functions?

ZNF367 (Zinc Finger Protein 367) is a transcription factor that functions as a central node in gene co-regulation networks during aging. Research has demonstrated that ZNF367 acts as a key controller of neuroblast cell cycle, particularly in the progression of mitosis and spindle checkpoint. The high conservation of ZNF367 across species, with 66% identity between human and Xenopus amino acid sequences (reaching 98% at zinc finger domains), suggests an evolutionarily conserved function in vertebrates from fish to primates . Additionally, ZNF367 has been identified as a potential oncogenic factor through its transcriptional activation of KIF15, which accelerates progression in certain cancers .

How is ZNF367 expressed during development?

ZNF367 is maternally expressed in the animal pole during Xenopus embryonic development. It shows specific expression patterns in neural tissue, including the eye field and neural crest cells. The spatial expression pattern of ZNF367 suggests a critical role in primary neurogenesis. This is supported by experimental evidence showing that knockdown of ZNF367 results in marked loss of post-mitotic neurons, indicating its essential function in neuronal differentiation . The gene appears to be involved in maintaining the appropriate balance between neural progenitor self-renewal and differentiation.

What types of ZNF367 antibodies are available for research?

Several types of ZNF367 antibodies are available for research applications, including:

• Rabbit polyclonal antibodies targeting the C-terminal region

• FITC-conjugated antibodies

• Biotin-conjugated antibodies

• HRP-conjugated antibodies

These antibodies have demonstrated reactivity across multiple species including human, cow, dog, horse, mouse, rabbit, rat, zebrafish, guinea pig, and pig . Most commercially available ZNF367 antibodies are validated for Western blotting applications, while some are also suitable for ELISA techniques. The diversity of available antibody formats allows researchers to select the most appropriate tool based on their specific experimental requirements.

What are the validated applications for ZNF367 antibodies?

ZNF367 antibodies have been primarily validated for Western blotting applications, which is critical for detecting and quantifying ZNF367 protein expression in various cell and tissue samples . Some antibodies are also validated for ELISA techniques. When designing experiments, researchers should consider that most commercially available antibodies target specific regions of the protein, such as the C-terminal region or amino acids 83-151 or 299-327. The application notes generally suggest that optimal working dilutions should be determined experimentally by each investigator to ensure maximum specificity and sensitivity .

What experimental controls should be included when using ZNF367 antibodies?

When using ZNF367 antibodies, researchers should implement several controls to ensure experimental validity:

• Positive controls: Include samples known to express ZNF367 (based on literature)

• Negative controls: Include samples where ZNF367 is absent or knocked down

• Loading controls: Use housekeeping proteins like β-tubulin (as demonstrated in the breast cancer studies) to normalize protein loading

• Isotype controls: Include appropriate isotype controls to detect non-specific binding

• Peptide competition assays: To confirm antibody specificity by pre-incubating with immunizing peptide

These controls help validate antibody specificity and ensure that observed signals truly represent ZNF367 protein detection rather than experimental artifacts or cross-reactivity with other proteins.

How can ZNF367 antibodies be used in neurogenesis research?

For neurogenesis research, ZNF367 antibodies can be employed through several methodological approaches:

• Immunohistochemistry: To visualize ZNF367 expression patterns in neural tissues during development

• Western blotting: To quantify ZNF367 protein levels during different stages of neuronal differentiation

• Co-immunoprecipitation: To identify protein interaction partners of ZNF367 in neural precursor cells

• ChIP-seq (Chromatin Immunoprecipitation followed by sequencing): To identify genomic binding sites of ZNF367 and its target genes in neural tissues

These applications can help elucidate ZNF367's role in maintaining the balance between neural progenitor self-renewal and differentiation. Research has shown that ZNF367 is required for neuronal differentiation but not for neuronal specification, as evidenced by unaffected expression of ngnr1 (a proneural marker) in ZNF367 morphants .

What is known about ZNF367's role in cell cycle regulation?

ZNF367 plays a critical role in cell cycle regulation, particularly during mitosis. Loss-of-function studies in Xenopus embryos revealed that ZNF367 knockdown results in:

• Increased expression of cell cycle markers (pcna)

• Higher number of mitotically active cells (measured by phosphorylated H3)

• Elevated expression of cyclin B1, which is predominantly expressed during M phase

These findings suggest that ZNF367 is required for proper exit from M phase or for control of the mitotic checkpoint preceding anaphase. In-silico analysis identified ZNF367 as a hub gene in a conserved module containing genes involved in cell cycle regulation, particularly those related to mitosis progression and spindle checkpoint . This functional role appears to be conserved across species and has implications for both developmental processes and pathological conditions like cancer.

How does ZNF367 relate to cancer progression mechanisms?

ZNF367 has been implicated in cancer progression through its transcriptional activation of KIF15. Research has shown that:

• ZNF367 acts as an upstream transcription factor for KIF15

• KIF15 is highly expressed in breast cancer tissues

• High KIF15 expression correlates with poor survival rates in breast cancer patients

• Silencing ZNF367 suppresses breast cancer cell viability, migration, invasion, and disrupts cell cycle distribution

• Overexpression of KIF15 can counteract the inhibitory effects of ZNF367 silencing on breast cancer progression

These findings suggest that the ZNF367-KIF15 axis represents a potential therapeutic target for cancer treatment. The relationship between ZNF367 and cell cycle regulation appears to be a common mechanism in both developmental contexts and cancer pathology.

What are the consequences of ZNF367 knockdown in neural development?

ZNF367 knockdown in Xenopus embryos leads to several significant phenotypic consequences:

• Strong reduction of cells expressing post-mitotic neuron markers (N-tubulin and elrC/HuC)

• Expanded expression domains of stemness genes (sox2 and rx1)

• Increased population of neural progenitors

• Enhanced self-renewal of progenitors at the expense of differentiation

• No significant increase in apoptosis

These observations suggest that ZNF367 is essential for the transition from proliferating neural progenitors to differentiated neurons. The loss of ZNF367 appears to disrupt this balance, leading to an accumulation of progenitors that fail to properly differentiate into mature neurons.

What genomic approaches can be used to identify ZNF367 target genes?

Several genomic approaches can be employed to identify ZNF367 target genes:

• ChIP-seq: To identify genomic regions directly bound by ZNF367

• RNA-seq following ZNF367 knockdown or overexpression: To identify genes whose expression changes in response to altered ZNF367 levels

• Weighted gene co-expression network analysis: This approach was successfully used to identify potential ZNF367 targets, revealing enrichment in genes involved in mitosis progression and spindle checkpoint

• Luciferase reporter assays: To validate direct transcriptional regulation, as demonstrated for KIF15 promoter activity by ZNF367

Through these approaches, researchers have identified potential ZNF367 targets including fancd2, ska3, and smc2. Expression domain analysis in Xenopus embryos confirmed expanded expression of these genes following ZNF367 knockdown .

How can protein-protein interactions of ZNF367 be studied?

Investigating ZNF367 protein-protein interactions requires sophisticated methodological approaches:

• Co-immunoprecipitation (Co-IP): Using ZNF367 antibodies to pull down protein complexes, followed by mass spectrometry analysis to identify interaction partners

• Proximity labeling techniques (BioID or APEX): Fusing ZNF367 to a biotin ligase to identify proteins in close proximity

• Yeast two-hybrid screening: To identify direct protein-protein interactions

• FRET (Fluorescence Resonance Energy Transfer) or BiFC (Bimolecular Fluorescence Complementation): To visualize protein interactions in living cells

These techniques can help elucidate the molecular complexes formed by ZNF367 during its function as a transcription factor and cell cycle regulator. Understanding these interaction networks is crucial for deciphering the mechanisms by which ZNF367 controls cellular processes.

What are the technical considerations for ZNF367 antibody storage and handling?

Proper storage and handling of ZNF367 antibodies is critical for maintaining their functionality:

• Storage temperature: Store at -20°C for long-term storage

• Short-term storage: 2-8°C for up to one week

• Aliquoting: Divide into small aliquots to prevent freeze-thaw cycles

• Buffer composition: Typically provided in 1x PBS buffer with 0.09% sodium azide and 2% sucrose

• Safety precautions: Contains sodium azide, which is hazardous and should be handled by trained staff only

Researchers should avoid repeated freeze-thaw cycles as this can degrade antibody quality and reduce specificity and sensitivity. Following manufacturer guidelines for specific antibody formats (unconjugated, FITC-conjugated, biotin-conjugated) is essential for optimal experimental results.

How does ZNF367 function relate to aging processes?

ZNF367 has been identified as an age-regulated gene that represents a central node in gene co-regulation networks during aging. Global population aging is a major social and economic challenge, and the progressive decline in physiological functions affects all organs, including the brain. The age-related incidence of neurodegenerative diseases coincides with the decline of adult neural stem cells. ZNF367's expression decreases with age, which correlates with its role in embryonic neurogenesis . This suggests that ZNF367 may be involved in the maintenance and repair of neuronal function throughout life, opening new avenues for investigation in adult neurogenesis and potential therapeutic approaches for age-related neurodegenerative conditions.

What experimental model systems are most appropriate for ZNF367 research?

Based on current research, several model systems have proven valuable for ZNF367 research:

• Xenopus laevis: Extensively used for developmental studies, allowing targeted knockdown in the CNS without affecting other tissues. The conservation of ZNF367 between human and Xenopus (66% identity at amino acid level) makes this a relevant model for functional studies .

• Human cell lines: Particularly cancer cell lines like MDA-MB-231 and SKBR3 have been used to study ZNF367's role in cancer progression .

• Neural stem/progenitor cells: Appropriate for studying ZNF367's role in neurogenesis and cell cycle regulation.

The choice of model system should be guided by the specific research question, with Xenopus being particularly useful for developmental studies and human cell lines for cancer-related research or for analyzing human-specific functions of ZNF367.