ZNF155 Antibody

What is ZNF155 and what are its key biological functions?

ZNF155 (Zinc finger protein 155) is a member of the krueppel C2H2-type zinc-finger protein family primarily localized in the nucleus. With a molecular weight of approximately 62.08 kDa in humans, ZNF155 functions as a transcriptional regulator involved in gene expression control and cell growth regulation . The protein is encoded by a gene located on chromosome 19q13.31 . ZNF155 contains characteristic zinc finger domains that enable sequence-specific DNA binding, allowing it to regulate the expression of target genes. Its involvement in transcriptional regulation makes it a subject of interest in developmental biology and cancer research . The protein has been studied in comparative genomics contexts, with research highlighting its presence in homologous gene clusters between humans and mice .

What types of ZNF155 antibodies are currently available for research?

Several types of ZNF155 antibodies are available to researchers, each with distinct characteristics suitable for various experimental applications:

The PCRP-ZNF155-1A5 monoclonal antibody was developed using recombinant protein as the immunogen and deposited to the Developmental Studies Hybridoma Bank by the Common Fund – Protein Capture Reagents Program . The rabbit polyclonal antibody was similarly raised against recombinant ZNF155 . Multiple antibody options provide researchers flexibility in experimental design based on their specific technical requirements and research questions .

How should ZNF155 antibodies be stored and handled to maintain optimal activity?

Upon receipt, researchers should verify the antibody's condition, aliquot appropriately based on experimental needs, and maintain proper documentation of storage conditions and freeze-thaw history. These handling practices are essential for ensuring reproducible experimental results and extending the antibody's useful lifespan.

What are the validated applications for ZNF155 antibodies in research?

ZNF155 antibodies have been validated for several research applications, with specific validations varying by antibody clone. The PCRP-ZNF155-1A5 monoclonal antibody is recommended for immunoprecipitation and microarray applications . Immunoprecipitation allows researchers to isolate ZNF155 protein complexes from cell lysates to identify interaction partners or study post-translational modifications. Microarray applications enable high-throughput screening of ZNF155 binding to various targets or assessment of expression patterns across different tissues or experimental conditions.

When selecting a ZNF155 antibody for research, it is crucial to consider the specific validation data provided by manufacturers. Validation typically includes confirmation of species reactivity, with the PCRP-ZNF155-1A5 antibody specifically validated for human ZNF155 . This species-specific validation is essential for ensuring experimental reliability and data interpretation accuracy when working with human samples or model systems.

How does one design proper controls for ZNF155 antibody experiments?

Designing appropriate controls is essential for reliable interpretation of ZNF155 antibody experiments. For basic experimental validation, include the following controls:

• Positive control: Use tissue or cell lines known to express ZNF155, such as those validated by antibody manufacturers through specificity testing . This confirms that the antibody can detect its intended target under your experimental conditions.

• Negative control: Include tissues or cells known to lack ZNF155 expression to verify specificity and assess background signal levels .

• Isotype control: For applications like immunohistochemistry or flow cytometry, include an isotype-matched antibody (e.g., MIgG2a for PCRP-ZNF155-1A5) that has no specific target in your sample to evaluate non-specific binding .

• Blocking peptide control: When available, pre-incubation of the antibody with a specific blocking peptide corresponding to the immunogen can demonstrate binding specificity.

• Knockdown/knockout validation: For definitive specificity confirmation, compare antibody signals between wild-type samples and those where ZNF155 has been depleted through genetic approaches.

These controls help distinguish between specific signal and experimental artifacts, ensuring confident interpretation of experimental results.

What is known about the specificity profile of commercially available ZNF155 antibodies?

The specificity of ZNF155 antibodies is a critical consideration for experimental reliability. Manufacturers typically validate antibody specificity through testing on tissues or cell samples with known positive and negative expression of ZNF155 . For the PCRP-ZNF155-1A5 monoclonal antibody, specificity for human ZNF155 has been confirmed, making it suitable for research using human samples .

Modern antibody development incorporates sophisticated techniques to ensure high specificity. Recent advances in antibody specificity inference and design utilize high-throughput sequencing and computational analysis to improve discrimination between very similar epitopes . This approach involves identifying distinct binding modes associated with particular ligands and has been successfully applied to disentangle binding patterns even between chemically similar targets .

When evaluating ZNF155 antibody specificity for research applications, consider:

• Cross-reactivity testing documentation provided by the manufacturer

• Validation in multiple applications to confirm consistent target recognition

• Literature citations demonstrating successful use in your intended application

• Epitope information, when available, to assess potential cross-reactivity with related zinc finger proteins

How can ZNF155 antibodies be employed in chromatin immunoprecipitation (ChIP) experiments to identify DNA binding sites?

For ChIP experiments investigating ZNF155 binding sites, a comprehensive approach is necessary due to the protein's role as a transcriptional regulator. Begin by optimizing crosslinking conditions; ZNF155 typically requires standard formaldehyde crosslinking (1% for 10 minutes at room temperature), but optimization may be necessary based on your specific cell type. Select a validated ChIP-grade ZNF155 antibody, with preference given to monoclonal antibodies that have demonstrated high specificity in immunoprecipitation applications .

The experimental protocol should include:

• Sonication optimization: Adjust sonication parameters to generate DNA fragments of 200-500 bp, verified by agarose gel electrophoresis prior to immunoprecipitation.

• Input controls: Reserve 5-10% of chromatin before immunoprecipitation to normalize for ChIP efficiency and starting material variations.

• Antibody validation: Perform Western blot analysis on input samples to confirm ZNF155 protein presence at expected molecular weight (approximately 62 kDa) .

• Multiple biological replicates: Conduct at least three independent biological replicates to ensure reproducibility of binding sites.

• Sequential ChIP: For investigating co-regulatory complexes, consider sequential ChIP with antibodies against known transcriptional cofactors.

For data analysis, use appropriate peak-calling algorithms (e.g., MACS2) and motif analysis tools to identify enriched DNA binding motifs. Compare results with published C2H2 zinc finger binding motifs to contextualize findings within the broader zinc finger protein family .

What approaches should be used to validate ZNF155 antibody specificity in complex experimental systems?

Validating ZNF155 antibody specificity in complex systems requires a multi-faceted approach:

• Genetic validation: Implement CRISPR/Cas9-mediated knockout or siRNA-mediated knockdown of ZNF155, followed by immunoblotting or immunostaining to confirm signal reduction or elimination. This provides the strongest evidence for antibody specificity.

• Recombinant protein competition assay: Pre-incubate the antibody with purified recombinant ZNF155 protein before application to samples. Specific antibody binding should be competitively inhibited, resulting in signal reduction.

• Peptide array analysis: Test antibody binding against peptide arrays containing ZNF155 sequences and related zinc finger protein sequences to assess cross-reactivity potential with homologous proteins.

• Mass spectrometry validation: Perform immunoprecipitation followed by mass spectrometry analysis to identify all proteins captured by the antibody. ZNF155 should be among the most abundant identified proteins.

• Orthogonal antibody comparison: Compare results using multiple ZNF155 antibodies with different epitopes. Consistent patterns across different antibodies provide stronger evidence for specific target detection.

Recent advances in antibody specificity inference utilize biophysics-informed models trained on phage display experimental data to predict and enhance antibody specificity profiles . These approaches can identify distinct binding modes associated with specific ligands, enabling the design and selection of antibodies with customized specificity profiles even for closely related targets .

How can ZNF155 antibodies be utilized in multiplex imaging to investigate protein-protein interactions?

Multiplex imaging techniques using ZNF155 antibodies require careful experimental design to identify potential protein-protein interactions within their native cellular context:

• Antibody compatibility assessment: Prior to multiplex experiments, test each antibody individually to confirm specific labeling and determine optimal dilutions. Select antibodies from different host species (e.g., PCRP-ZNF155-1A5 from mouse and polyclonal antibodies from rabbit) to enable simultaneous detection with species-specific secondary antibodies .

• Proximity ligation assay (PLA): This technique can detect ZNF155 interactions with suspected binding partners at single-molecule resolution. PLA requires antibodies against both proteins of interest, followed by species-specific secondary antibodies conjugated to oligonucleotides that enable rolling circle amplification when proteins are in close proximity (<40 nm).

• Advanced microscopy approaches:

  • STORM/PALM super-resolution microscopy: Enables visualization of ZNF155 colocalization with potential interaction partners at nanoscale resolution

  • FRET microscopy: When using fluorescently labeled antibodies, can demonstrate direct protein-protein interactions through energy transfer

  • Multiplexed ion beam imaging (MIBI): Allows simultaneous detection of multiple proteins using metal-labeled antibodies

• Sequential immunostaining protocols: For investigating multiple markers using antibodies from the same species, implement sequential staining with intermediary stripping or quenching steps. This approach is particularly valuable for tissues with limited availability.

• Computational analysis: Apply machine learning algorithms to quantify colocalization patterns and identify statistically significant associations between ZNF155 and other nuclear proteins across multiple cells and experimental conditions.

These approaches enable researchers to move beyond simple colocalization to more definitively establish functional protein-protein interactions involving ZNF155 in their physiological context.

What are common troubleshooting strategies for non-specific binding with ZNF155 antibodies?

When encountering non-specific binding with ZNF155 antibodies, systematically address the issue through these methodological approaches:

• Blocking optimization: Test different blocking agents (BSA, normal serum, casein, or commercial blocking solutions) at various concentrations and incubation times. For ZNF155 detection in nuclear extracts, use 5% non-fat dry milk in TBST for 1-2 hours at room temperature as a starting point.

• Antibody dilution titration: Perform serial dilutions of the primary antibody to determine the optimal concentration that maximizes specific signal while minimizing background. For Western blots, begin with a 1:1000 dilution of the PCRP-ZNF155-1A5 antibody and adjust as needed .

• Washing protocol refinement: Increase washing stringency by:

  • Adding 0.1-0.3% Triton X-100 to wash buffers

  • Extending wash durations (5-10 minutes per wash)

  • Increasing the number of wash steps (5-6 washes)

  • Using high-salt wash buffers (up to 500 mM NaCl) for particularly problematic non-specific binding

• Pre-adsorption techniques: For tissue immunohistochemistry, pre-adsorb the antibody with tissues known to lack ZNF155 expression to remove antibodies with non-specific binding potential.

• Sample preparation optimization: For nuclear proteins like ZNF155, ensure complete nuclear lysis and consider using specialized nuclear extraction buffers containing DNase to release DNA-bound proteins.

• Secondary antibody considerations: Test alternative secondary antibodies from different manufacturers, and consider using highly cross-adsorbed secondary antibodies to minimize species cross-reactivity.

For Western blot applications specifically, cutting membranes to include only the region around the expected 62 kDa size of ZNF155 can help eliminate visualization of non-specific bands at other molecular weights .

How should researchers optimize ZNF155 antibody concentrations for different experimental applications?

Optimization of ZNF155 antibody concentrations should be tailored to each experimental application:

For each application, perform preliminary experiments with positive and negative controls. For Western blot optimization, include lysates from cells known to express ZNF155 alongside negative controls. For immunostaining applications, include tissues or cells with validated ZNF155 expression patterns. Document all optimization steps methodically, as different lots of the same antibody may require re-optimization .

What considerations should be taken into account when using ZNF155 antibodies across different species?

When applying ZNF155 antibodies across different species, several critical factors must be considered:

• Validated species reactivity: The PCRP-ZNF155-1A5 monoclonal antibody has been specifically validated for human ZNF155, while reactivity with other species must be empirically determined . Always verify the manufacturer's documentation regarding species cross-reactivity before proceeding.

• Sequence homology analysis: Prior to experimental application, perform sequence alignment of the ZNF155 protein across species of interest, focusing particularly on the epitope region if known. Higher sequence conservation increases the likelihood of cross-reactivity.

• Evolutionary considerations: Zinc finger proteins like ZNF155 often undergo differential expansion in homologous gene clusters between species. Research has specifically documented differences between human and mouse zinc finger gene clusters , suggesting potential challenges in antibody cross-reactivity.

• Pilot studies requirement: When using ZNF155 antibodies in non-validated species, conduct preliminary validation experiments including:

  • Western blot analysis to confirm band size and specificity

  • Immunoprecipitation followed by mass spectrometry to verify target identity

  • Side-by-side comparison with positive control samples from validated species

• Specificity controls: Include more rigorous specificity controls when working with non-validated species, such as competitive blocking with recombinant protein and genetic knockdown approaches.

• Interpretation caution: Exercise particular caution when interpreting negative results in non-validated species, as lack of signal could indicate antibody incompatibility rather than absence of the protein.

For cross-species applications, polyclonal antibodies may offer advantages in some cases due to their recognition of multiple epitopes, potentially increasing the chance of cross-reactivity with homologous proteins across species .

How can computational approaches enhance ZNF155 antibody specificity for research applications?

Recent advances in computational biology have revolutionized antibody specificity optimization, with significant implications for ZNF155 research:

• Biophysics-informed modeling: State-of-the-art approaches combine high-throughput sequencing with biophysical modeling to predict and design antibody specificity beyond experimentally observed sequences . These models associate distinct binding modes with particular ligands, allowing for the computational disentanglement of binding patterns even between chemically similar epitopes .

• Machine learning applications: Supervised machine learning algorithms trained on phage display experimental data can predict antibody-epitope interactions and identify key residues that determine specificity . For ZNF155 antibodies, this approach could help:

  • Identify antibody variants with enhanced specificity for ZNF155 over related zinc finger proteins

  • Design antibodies that recognize specific post-translational modifications of ZNF155

  • Create antibodies that distinguish between potential ZNF155 isoforms

• In silico epitope mapping: Computational tools can predict conformational and linear epitopes on ZNF155, guiding antibody selection for specific experimental needs. This is particularly valuable for distinguishing between functional domains of the protein.

• Counter-selection strategies: Computational approaches can efficiently achieve counter-selection to eliminate off-target binding, a process more challenging to accomplish experimentally . These methods can generate antibody variants with customized specificity profiles, either with specific high affinity for ZNF155 or with controlled cross-specificity for multiple related targets .

• Implementation methodology: Researchers can collaborate with computational biology specialists to apply these approaches, or utilize commercial services that incorporate these computational methods in custom antibody development. Several academic platforms also provide tools for antibody sequence optimization based on specificity parameters.

These computational approaches offer significant advantages for developing highly specific ZNF155 antibodies, particularly for discriminating between closely related zinc finger family members that may share structural similarities .

What emerging technologies are expanding the applications of ZNF155 antibodies in single-cell analysis?

Single-cell analysis technologies are rapidly advancing the utility of ZNF155 antibodies for understanding cellular heterogeneity:

• Single-cell proteomics approaches:

  • Mass cytometry (CyTOF): Utilizes metal-tagged antibodies, including those against ZNF155, to simultaneously quantify dozens of proteins at single-cell resolution. This approach can reveal rare cell populations with distinct ZNF155 expression patterns.

  • Cellular indexing of transcriptomes and epitopes by sequencing (CITE-seq): Combines antibody detection with transcriptome analysis, allowing correlation between ZNF155 protein levels and gene expression profiles in individual cells.

• Spatial proteomics technologies:

  • Imaging mass cytometry: Provides spatial context for ZNF155 expression relative to other proteins in tissue sections at subcellular resolution.

  • Co-detection by indexing (CODEX): Enables highly multiplexed imaging using DNA-barcoded antibodies, including ZNF155 antibodies, with iterative imaging cycles.

  • Spatial transcriptomics combined with immunofluorescence: Correlates ZNF155 protein localization with local transcriptional landscapes.

• Methodological considerations:

  • Antibody conjugation protocols must be optimized for each platform to maintain specificity and sensitivity

  • Signal amplification strategies may be necessary for detecting low-abundance ZNF155 in certain cell types

  • Computational analysis pipelines must account for technical variation and background signals

• Validation requirements:

  • Single-cell technologies require rigorous validation of antibody specificity

  • Orthogonal validation approaches (e.g., fluorescence microscopy, flow cytometry) should confirm single-cell findings

  • Genetic perturbation experiments should verify antibody specificity at single-cell resolution

These technologies enable unprecedented insights into ZNF155 function in heterogeneous cell populations, potentially revealing cell type-specific roles in development, disease progression, and response to therapeutic interventions.

What are the emerging trends in ZNF155 antibody development and application?

The field of ZNF155 antibody development is evolving rapidly, with several key trends emerging that promise to enhance research capabilities:

• Integration of computational and experimental approaches: The combination of biophysics-informed modeling with extensive selection experiments is creating antibodies with precisely tailored specificity profiles . This convergence allows researchers to design ZNF155 antibodies that can either specifically target ZNF155 or recognize defined subsets of related zinc finger proteins, depending on research requirements.

• Domain-specific antibodies: Development is progressing toward antibodies that recognize specific functional domains of ZNF155, such as the KRAB A domain versus the zinc finger domains . This domain-specific approach enables more nuanced investigations of protein function.

• Expansion to modification-specific antibodies: There is growing interest in antibodies that specifically recognize post-translationally modified forms of ZNF155, allowing researchers to investigate regulatory mechanisms controlling ZNF155 function.

• Application in advanced imaging technologies: ZNF155 antibodies are increasingly being adapted for use in super-resolution microscopy and highly multiplexed imaging approaches, enabling spatial analysis of ZNF155 in relation to other nuclear proteins and chromatin features.

• Incorporation into single-cell multi-omics: The integration of ZNF155 antibodies into single-cell multi-omic platforms is allowing researchers to correlate protein expression with transcriptomic and epigenomic features at unprecedented resolution.

These trends collectively point toward more precise, versatile tools for investigating ZNF155 biology in increasingly complex experimental systems, driving deeper understanding of its roles in health and disease.

How should researchers evaluate contradictory results obtained with different ZNF155 antibodies?

When faced with contradictory results using different ZNF155 antibodies, a systematic troubleshooting and validation approach is essential:

• Comprehensive antibody characterization:

  • Compare epitope specificity: Different antibodies may recognize distinct epitopes on ZNF155, potentially affected differently by protein conformation, interactions, or modifications

  • Review clonality differences: Results from monoclonal antibodies (e.g., PCRP-ZNF155-1A5) and polyclonal antibodies may vary due to their inherent properties

  • Examine species of origin: Host species can influence antibody performance in certain applications or with particular detection systems

• Technical validation experiments:

  • Perform side-by-side comparisons using standardized protocols and identical samples

  • Implement genetic validation through ZNF155 knockdown/knockout to determine which antibody most accurately reflects true protein levels

  • Utilize orthogonal detection methods (e.g., mass spectrometry) to verify results independently of antibody-based approaches

• Biological context considerations:

  • Evaluate potential isoform specificity: Contradictory results may reflect detection of different ZNF155 isoforms

  • Assess post-translational modification sensitivity: Some antibodies may be affected by phosphorylation, SUMOylation, or other modifications

  • Consider protein complex formation: Epitope masking in certain protein complexes may affect antibody accessibility

• Methodological approach to resolution:

  • Document all experimental conditions comprehensively

  • Test antibodies under varied fixation/extraction conditions

  • Consider employing tagged ZNF155 constructs as definitive controls

  • Consult literature and manufacturer technical support for known limitations

• Reporting and interpretation:

  • Clearly report contradictory findings in publications rather than selectively presenting consistent results

  • Discuss potential technical and biological explanations for differences

  • Consider the contradictions as potentially informative about ZNF155 biology rather than merely technical failures