znf687a Antibody

What is ZNF687 and why is it significant in cancer research?

ZNF687 (Zinc finger protein 687) functions as a central hub in a large transcriptional regulatory network. Initially identified as a translocation partner gene with RUNX1 in patients with acute myeloid leukemia (AML), ZNF687 has emerged as a protein of interest in multiple cancer types . Recent studies have demonstrated its oncogenic role in lung adenocarcinoma (LUAD), where elevated expression correlates with poorer prognosis, enhanced tumor growth, migration, invasion, and promotion of epithelial-mesenchymal transition (EMT) . Understanding ZNF687's molecular function is critical as it may be involved in chromatin-modifying complexes essential for embryonic development and stem cell renewal through its interaction with the Ring1/Rnf2 RING finger protein member of the Polycomb group .

What isoforms of ZNF687 exist and which ones can be detected by commercially available antibodies?

According to current research, at least three isoforms of ZNF687 are known to exist. Most commercially available antibodies, including those from Rockland, Abnova, and Boster Bio, will detect the two largest isoforms . This specificity is important for experimental design, particularly when investigating isoform-specific functions. Researchers should verify which isoforms their selected antibody targets before proceeding with experiments to ensure accurate interpretation of results.

What are the standard applications for ZNF687 antibodies in research settings?

ZNF687 antibodies have been validated for multiple research applications including:

Most ZNF687 antibodies are human-specific, and researchers should optimize conditions for their specific experimental setup . When planning experiments, consider that ZNF687 expression is elevated in certain cancer cell lines, including A549, HCC827, H1975, and H1299 lung adenocarcinoma cells, which may serve as positive controls .

How should I optimize ZNF687 antibody dilutions for immunohistochemistry applications?

For immunohistochemistry applications with ZNF687 antibodies, begin with the manufacturer's recommended dilution (typically around 5 μg/mL for paraffin-embedded sections) . To determine optimal dilution:

• Perform a titration experiment using serial dilutions (e.g., 2.5, 5, and 10 μg/mL)

• Include positive control tissues known to express ZNF687 (e.g., human tonsil tissue or LUAD tissue samples)

• Include negative controls (primary antibody omission and tissues with low ZNF687 expression like PC9, BEAS-2B, or HBE cell lines)

• Evaluate signal-to-noise ratio, specific staining patterns, and background

• Select the dilution that provides the highest signal-to-noise ratio while maintaining specificity

For paraffin sections, ensure proper antigen retrieval methods are employed as ZNF687 may require specific conditions for optimal epitope exposure .

What are the recommended storage conditions for maintaining ZNF687 antibody stability and performance?

To maintain optimal stability and performance of ZNF687 antibodies, follow these storage protocols:

For optimal results, aliquot the antibody solution into smaller volumes before freezing to minimize freeze-thaw cycles . Centrifuge the product if it is not completely clear after standing at room temperature. Most ZNF687 antibodies maintain stability for several weeks at 4°C as undiluted liquid but should be diluted only immediately before use .

What positive and negative controls should I use when validating ZNF687 antibody specificity?

When validating ZNF687 antibody specificity, appropriate controls are essential:

Recommended Positive Controls:

• Cell lines: A549, HCC827, H1975, and H1299 (high ZNF687 expression)

• Tissues: Human tonsil tissue, LUAD tissues

Recommended Negative Controls:

• Cell lines: PC9, BEAS-2B, and HBE (low ZNF687 expression)

• Primary antibody omission control

• Blocking peptide competition assay using the immunogen peptide

• Tissues from control subjects without disease (e.g., paracarcinoma tissues for cancer studies)

For advanced validation, consider using ZNF687 knockdown cells alongside overexpression models to confirm antibody specificity through differential signal intensity .

How can ZNF687 antibodies be used to investigate the role of ZNF687 in cancer progression pathways?

ZNF687 antibodies can be employed in multiple sophisticated approaches to investigate its role in cancer progression:

• Chromatin Immunoprecipitation (ChIP) assays:

  • Use ZNF687 antibodies to identify genomic binding sites and target genes

  • Combine with sequencing (ChIP-seq) to create genome-wide binding profiles

  • Correlate binding patterns with transcriptional changes

• Co-immunoprecipitation (Co-IP) studies:

  • Investigate protein-protein interactions between ZNF687 and:

    • Components of the PI3K/AKT signaling pathway

    • ZNF592 and ZMYMD8 in the proposed transcriptional network

    • Ring1/Rnf2 and other Polycomb group proteins

• Signaling pathway analysis:

  • Combine with phospho-specific antibodies to monitor PI3K/AKT pathway activation

  • Study how ZNF687 affects cell cycle regulators (p21, p27, p53, CDK2, CDK4, CDK6, cyclin D1, and cyclin D3)

• EMT marker correlation:

  • Perform multiplex immunofluorescence to simultaneously detect ZNF687 and EMT markers

  • Analyze tissue microarrays to correlate ZNF687 expression with clinical outcomes

These approaches can help elucidate the mechanistic role of ZNF687 in cancer progression and potentially identify novel therapeutic targets.

What methodological approaches can resolve conflicting ZNF687 antibody results in different experimental systems?

When encountering conflicting ZNF687 antibody results across different experimental systems, implement these troubleshooting strategies:

• Antibody validation with multiple technical approaches:

  • Confirm specificity using at least two independent detection methods (e.g., IHC, Western blot, ELISA)

  • Employ antibodies targeting different epitopes of ZNF687

  • Verify results with siRNA/shRNA knockdown experiments

• Cell line and tissue verification:

  • Consider cell line authentication to eliminate potential misidentification

  • Sequence the ZNF687 gene in your experimental system to check for mutations or variants

  • Evaluate tissue-specific post-translational modifications that might affect antibody binding

• Protocol optimization for specific experimental systems:

  • Adjust fixation methods, as over-fixation can mask epitopes

  • Test different antigen retrieval methods for IHC applications

  • Optimize blocking and permeabilization conditions for different cell types

• Cross-reactivity assessment:

  • Use tissues/cells from ZNF687 knockout models as negative controls

  • Test antibody specificity in systems with known ZNF687 expression profiles

  • Evaluate potential cross-reactivity with other zinc finger proteins with similar domains

These methodological approaches can help resolve discrepancies and ensure reliable, reproducible results across different experimental systems.

How can ZNF687 antibodies be used to investigate the relationship between ZNF687 and cell cycle regulation in cancer models?

To investigate ZNF687's role in cell cycle regulation using antibodies:

• Flow cytometry with dual staining:

  • Combine ZNF687 antibody staining with propidium iodide or DAPI for cell cycle analysis

  • Correlate ZNF687 expression levels with cell cycle phase distribution

  • Use ZNF687 overexpression and knockdown models to observe cell cycle perturbations

• Cell cycle protein interaction studies:

  • Perform co-immunoprecipitation with ZNF687 antibodies followed by mass spectrometry

  • Investigate interactions with key cell cycle regulators:

    • p21, p27, p53 (cell cycle inhibitors)

    • CDK2, CDK4, CDK6 (cyclin-dependent kinases)

    • Cyclins D1 and D3

• Chromatin association dynamics:

  • Use chromatin fractionation followed by immunoblotting with ZNF687 antibodies

  • Analyze ZNF687's chromatin association patterns across cell cycle phases

  • Combine with synchronization protocols to capture specific cell cycle stages

• Transcriptional regulation analysis:

  • Conduct ChIP-seq with ZNF687 antibodies at different cell cycle phases

  • Identify cell cycle-dependent binding to promoters of cell cycle genes

  • Correlate with RNA-seq data to establish functional consequences

Research has shown that ZNF687 knockdown increases the proportion of cancer cells in G1 phase, while overexpression enhances G1-S transition, suggesting a direct involvement in cell cycle progression that can be further explored using these approaches .

What are the critical factors affecting reproducibility when using ZNF687 antibodies in different experimental contexts?

Several factors can significantly impact reproducibility when working with ZNF687 antibodies:

• Antibody source and lot variation:

  • Different lots may show varying performance characteristics

  • Maintain detailed records of antibody lots used

  • Consider antibody validation with each new lot

• Sample preparation variability:

  • Standardize fixation protocols (duration, temperature, fixative composition)

  • Control epitope masking through consistent antigen retrieval methods

  • Ensure consistent cell lysis conditions for protein extraction

• Detection system differences:

  • Standardize secondary antibody selection and dilution

  • Use consistent visualization methods (fluorescent vs. enzymatic)

  • Control for autofluorescence in fluorescence-based applications

• ZNF687 expression fluctuations:

  • Account for cell density effects on expression

  • Control for cell cycle phase as ZNF687 levels may vary during cell cycle progression

  • Consider potential induction by cellular stress responses

• Post-translational modifications:

  • Be aware that phosphorylation or other modifications may affect antibody binding

  • Use phosphatase inhibitors during protein extraction if phosphorylation is suspected

  • Consider modification-specific antibodies for comprehensive analysis

Implementing rigorous standardization protocols for these factors can significantly improve reproducibility across experiments and between laboratories.

How can ZNF687 antibodies be incorporated into multiplexed immunoassays to study cancer signaling networks?

Incorporating ZNF687 antibodies into multiplexed immunoassays requires strategic planning:

• Panel design considerations:

  • Select antibodies raised in different host species to avoid cross-reactivity

  • Choose fluorophores with minimal spectral overlap for immunofluorescence

  • Include ZNF687 alongside PI3K/AKT pathway components based on established connections

• Optimization strategies:

  • Titrate each antibody individually before combining in multiplexed format

  • Perform sequential staining with intermediate blocking steps if cross-reactivity occurs

  • Validate multiplex results against single-plex controls

• Advanced multiplexing technologies:

  • Mass cytometry (CyTOF) using metal-conjugated ZNF687 antibodies

  • Multiplex immunohistochemistry with tyramide signal amplification

  • Proximity ligation assays to study ZNF687 protein-protein interactions

• Data analysis approaches:

  • Employ dimensionality reduction techniques (tSNE, UMAP) for high-parameter data

  • Utilize hierarchical clustering to identify co-expression patterns

  • Apply machine learning algorithms to correlate ZNF687 with other cancer markers

A recommended multiplexed panel for studying ZNF687 in cancer contexts might include:

This approach enables simultaneous assessment of ZNF687 expression in relation to key signaling and phenotypic markers in cancer research .

What considerations should be made when selecting between polyclonal and monoclonal ZNF687 antibodies for specific research applications?

When choosing between polyclonal and monoclonal ZNF687 antibodies, consider these application-specific factors:

Polyclonal ZNF687 Antibodies:

• Advantages:

  • Recognize multiple epitopes, providing robust detection even if some epitopes are masked

  • Higher sensitivity for low-abundance ZNF687 detection

  • Better performance in applications like immunoprecipitation and ChIP

  • More tolerance to minor protein denaturation or modifications

• Best applications:

  • Initial characterization studies

  • Immunoprecipitation of ZNF687 and associated complexes

  • ChIP experiments to study ZNF687 genomic binding

  • Detection of denatured ZNF687 in western blotting

Monoclonal ZNF687 Antibodies:

• Advantages:

  • Higher specificity for a single epitope

  • Reduced background and cross-reactivity

  • Lower lot-to-lot variability

  • Better suited for quantitative applications

• Best applications:

  • Flow cytometry analysis of ZNF687

  • Super-resolution microscopy

  • Quantitative western blotting

  • Standardized diagnostic applications

Application-Specific Selection Guidelines: