ZNF816 Antibody

What is ZNF816 and what is its biological function?

ZNF816 (also known as ZNF816A or Zinc finger protein 816) is a protein involved in transcriptional regulation. It functions as part of a zinc finger protein complex that coordinates DNA-binding activities necessary for gene expression. ZNF816A is expressed in various tissues with notable levels observed in the brain and immune cells . The protein may be involved in regulating the expression of target genes, though specific transcriptional targets and regulatory pathways remain under investigation.

What types of ZNF816 antibodies are currently available for research?

The research community has access to several ZNF816 antibody options with different applications and specifications:

How does ZNF816 expression vary across different tissue types?

ZNF816A shows a distinctive expression pattern across human tissues:

This expression pattern suggests ZNF816 may have specific roles in neural tissue, immune function, and potentially in certain cancer types.

What are the optimal conditions for using ZNF816 antibodies in immunohistochemistry?

For successful ZNF816 detection in paraffin-embedded tissues, researchers should consider the following protocol:

• Sample preparation: Standard deparaffinization and rehydration

• Antigen retrieval: Heat-induced epitope retrieval in citrate buffer (pH 6.0)

• Blocking: 5-10% normal serum from the same species as the secondary antibody

• Primary antibody: Dilute ZNF816 antibody at 1:100 dilution (optimal for ab234688)

• Secondary detection: Use appropriate species-specific detection system

• Visualization: DAB substrate for chromogenic detection or fluorescence imaging

• Controls: Include negative controls (primary antibody omission) and positive controls (tissues with known ZNF816 expression)

Published research has successfully demonstrated ZNF816 staining in liver cancer and colon cancer tissues using this approach .

What validation strategies should be employed when using ZNF816 antibodies?

Comprehensive validation of ZNF816 antibodies should include:

• Specificity testing:

  • Multiple antibodies targeting different epitopes

  • Knockdown/knockout controls (siRNA, CRISPR-Cas9)

  • Peptide competition assays

  • Western blot confirmation of specific band at predicted molecular weight

• Application-specific validation:

  • For IHC: Compare staining patterns across multiple tissue types

  • For ICC/IF: Confirm expected subcellular localization (primarily nuclear)

  • For WB: Verify band size matches predicted molecular weight

  • For all applications: Include appropriate positive and negative controls

• Cross-reactivity assessment:

  • Test in tissues/cells known to lack ZNF816 expression

  • Check for potential cross-reactivity with other zinc finger proteins

How can ZNF816 antibodies be incorporated into multi-parameter analyses?

When designing multi-parameter studies involving ZNF816:

• For immunofluorescence co-localization:

  • Select compatible primary antibodies (different host species)

  • Choose fluorophores with minimal spectral overlap

  • Optimize fixation and permeabilization for nuclear proteins

  • Perform sequential staining if antibody compatibility is limited

• For flow cytometry:

  • Use appropriate fixation/permeabilization buffers for nuclear proteins

  • Include proper compensation controls for each fluorophore

  • Use FMO (fluorescence minus one) controls

  • Incorporate lineage markers to identify specific cell populations

• For multiplexed IHC:

  • Consider sequential IHC with stripping or multispectral imaging approaches

  • Include single-stain controls for each antibody

  • Use automated image analysis software for quantification

How can ZNF816 antibodies be used to investigate its role in transcriptional regulation?

To elucidate ZNF816's function in transcriptional regulation:

• Chromatin Immunoprecipitation (ChIP):

  • Use ZNF816 antibodies to immunoprecipitate protein-DNA complexes

  • Perform ChIP-seq to identify genome-wide binding sites

  • Analyze binding motifs to determine DNA sequence preferences

  • Compare binding sites with genomic features (promoters, enhancers)

• Protein interaction studies:

  • Use co-immunoprecipitation (Co-IP) with ZNF816 antibodies

  • Identify protein interaction partners via mass spectrometry

  • Validate interactions using reciprocal Co-IP or proximity ligation assays

• Transcriptional analysis:

  • Modulate ZNF816 expression and perform RNA-seq

  • Correlate ZNF816 binding sites with expression changes

  • Use reporter gene assays to confirm direct regulation

What protocols are recommended for troubleshooting weak or non-specific ZNF816 antibody signals?

When facing technical challenges with ZNF816 antibody applications:

How can researchers integrate ZNF816 protein data with genomic and transcriptomic analyses?

For multi-omics integration involving ZNF816:

• Correlative approaches:

  • Compare ZNF816 protein expression with corresponding mRNA levels

  • Investigate discrepancies that might indicate post-transcriptional regulation

  • Correlate with genomic alterations (mutations, CNVs) affecting ZNF816

• Functional genomics:

  • Use ChIP-seq with ZNF816 antibodies to identify DNA binding sites

  • Integrate binding data with gene expression changes after ZNF816 modulation

  • Incorporate chromatin accessibility data (ATAC-seq) to assess binding context

• System-level analysis:

  • Construct gene regulatory networks incorporating ZNF816

  • Identify co-regulated genes and pathways

  • Use machine learning approaches to predict ZNF816 function based on integrated data

What methods are recommended for detecting ZNF816 in cancer tissues?

For optimal detection of ZNF816 in cancer samples:

• IHC protocol optimization:

  • Use 1:100 dilution of ZNF816 antibody (successful in liver and colon cancer)

  • Include appropriate positive controls (PC-3 cells or known positive tissues)

  • Compare with adjacent normal tissue when possible

• Cancer-specific considerations:

  • Assess tumor heterogeneity by examining multiple regions

  • Correlate with tumor grade, stage, and molecular subtypes

  • Consider double staining with proliferation or differentiation markers

• Quantification approaches:

  • Develop standardized scoring system for ZNF816 expression

  • Use digital pathology for objective quantification

  • Correlate expression with clinical parameters and outcomes

How should researchers interpret subcellular localization patterns of ZNF816?

When analyzing ZNF816 localization:

• Expected patterns:

  • Primary nuclear localization (consistent with transcription factor function)

  • Potential nucleolar or chromatin-associated distribution patterns

  • Possible cytoplasmic localization under specific conditions

• Analytical approaches:

  • High-resolution confocal microscopy for detailed nuclear distribution

  • Co-staining with nuclear subcompartment markers

  • Quantitative analysis of nuclear vs. cytoplasmic distribution

• Functional implications:

  • Changes in localization during cell cycle progression

  • Alterations in response to signaling events or stress

  • Correlation with transcriptional activity

What are the best practices for using ZNF816 antibodies in flow cytometry?

For flow cytometry applications with ZNF816 antibodies:

• Sample preparation:

  • Robust fixation (4% paraformaldehyde)

  • Effective nuclear permeabilization (methanol or commercial permeabilization buffers)

  • Single-cell suspension preparation

• Staining protocol:

  • Titrate antibody to determine optimal concentration

  • Include unstained, isotype, and FMO controls

  • Consider sequential staining if combining with other nuclear markers

• Analysis considerations:

  • Gate on single cells and viable populations

  • Set positive/negative thresholds using controls

  • Consider analyzing median fluorescence intensity rather than percent positive

How might ZNF816 antibodies contribute to understanding its potential role in neurobiology?

Given the notable expression of ZNF816 in brain tissue , researchers might:

• Investigate neuroanatomical distribution:

  • Map expression across brain regions using IHC with ZNF816 antibodies

  • Identify specific neuronal or glial populations expressing ZNF816

  • Analyze developmental expression patterns

• Explore functional implications:

  • Study ZNF816 expression in neurological disease models

  • Examine regulation during neural development and plasticity

  • Investigate target genes in neural tissues through ChIP-seq

• Apply in neurobiology research:

  • Use in co-localization studies with neural markers

  • Apply to patient-derived samples from neurological conditions

  • Incorporate into single-cell analysis of brain tissue

What considerations exist for developing custom ZNF816 antibodies for specialized research applications?

When developing custom ZNF816 antibodies:

• Epitope selection strategies:

  • Target unique regions outside the zinc finger domains to avoid cross-reactivity

  • Consider known post-translational modification sites

  • Design epitopes based on predicted surface exposure

  • Account for potential structural constraints

• Validation requirements:

  • Comprehensive specificity testing in multiple applications

  • Comparison with existing commercial antibodies

  • Validation in knockout/knockdown systems

  • Application-specific optimization

• Specialized applications:

  • Super-resolution microscopy compatible antibodies

  • Directly conjugated antibodies for multiplexed imaging

  • ChIP-grade antibodies with high specificity and efficiency

  • Cross-species reactive antibodies for comparative studies