znf503 Antibody

What is ZNF503 and what are its key biological functions?

ZNF503 (Zinc Finger Protein 503) is a transcriptional repressor involved in critical developmental processes and cellular differentiation. It contains zinc finger domains that enable DNA binding and modulation of specific target gene transcription . ZNF503 plays substantial roles in:

• Embryonic development, particularly in neural and mesenchymal tissues

• Retinal pigment epithelium (RPE) differentiation and optic fissure closure

• Regulation of stem cell differentiation

• Neuronal migration, immune response, and organogenesis

• Cancer progression through modulation of gene networks involved in cell proliferation and differentiation

What types of ZNF503 antibodies are available for research applications?

Several types of ZNF503 antibodies are available for research:

• Polyclonal antibodies raised against full-length human ZNF503 protein

• Polyclonal antibodies targeting specific amino acid regions (AA 1-646, AA 250-350, AA 100-300)

• Antibodies validated for various applications including Western blot, immunoprecipitation, immunohistochemistry, and immunofluorescence

• Host species typically include rabbit and mouse, with reactivity primarily against human ZNF503

Which experimental applications are most effective for ZNF503 detection and analysis?

The most effective applications for ZNF503 detection and analysis include:

The choice of application depends on the specific research question, with Western blot being particularly useful for expression level studies and immunohistochemistry for tissue-specific localization .

What are the optimal conditions for Western blot analysis of ZNF503?

For optimal Western blot detection of ZNF503:

• Use a concentration of 0.1-0.4 μg/mL of antibody

• Include appropriate positive controls such as 293T or MCF7 cell lysates

• Expect multiple potential bands (28 kDa, 55 kDa, 63 kDa) representing different isoforms

• Use ECL detection systems for optimal visualization

• For exposure time, start with 3 minutes and adjust based on signal strength

• Block with 5% non-fat milk or BSA in TBST

• Consider using PVDF membranes for better protein retention

How can I optimize immunohistochemical detection of ZNF503 in tissue samples?

For optimal immunohistochemical detection of ZNF503:

• Use a 1/200 dilution of validated antibodies (such as ab254715)

• Perform proper antigen retrieval (heat-induced epitope retrieval in citrate buffer pH 6.0)

• Include positive control tissues with known ZNF503 expression (endometrium, rectum, prostate have shown positive staining)

• Include negative control tissues like tonsil, which shows low positivity in lymphoid cells

• Use a detection system compatible with the host species of the primary antibody

• Counterstain appropriately to visualize tissue architecture

• Allow sufficient incubation time (overnight at 4°C for primary antibody often yields best results)

What is the normal expression pattern of ZNF503 in human tissues?

ZNF503 shows a dynamic expression pattern across human tissues:

• Expressed in endometrium, rectum, and prostate tissues

• Low expression in lymphoid cells of tonsil tissue

• Dynamically expressed during embryonic development, particularly in developing mouse eyes

• Expression is regulated by developmental signaling pathways including hedgehog and BMP signaling

• Found in multiple cell lines, with varying expression levels (higher in cancer cell lines like MCF7 and PC-3)

How is ZNF503 dysregulated in disease states, and what research applications explore this?

ZNF503 dysregulation has been documented in several disease states:

Research applications typically include expression analysis in patient samples, functional studies in cell lines, and animal models to determine causality .

How can ZNF503 antibodies be used to investigate transcriptional regulatory networks?

ZNF503 antibodies can be utilized to investigate transcriptional regulatory networks through:

• Chromatin Immunoprecipitation (ChIP) assays:

  • ZNF503 has been shown to directly bind the GATA3 promoter in hepatocellular carcinoma cells

  • ChIP followed by sequencing (ChIP-seq) can identify genome-wide binding sites

• Co-immunoprecipitation (Co-IP) studies:

  • Identify protein-protein interactions between ZNF503 and other transcriptional regulators

  • Coupled with mass spectrometry to discover novel interaction partners

• Sequential ChIP (Re-ChIP):

  • Determine if ZNF503 forms complexes with other transcription factors at specific genomic loci

• Transcription factor activity assays:

  • Using reporter gene constructs to measure the effect of ZNF503 on target promoter activity

  • For example, ZNF503 has been shown to repress GATA3 expression in HCC cells

These approaches can reveal how ZNF503 contributes to developmental processes and disease progression through transcriptional regulation .

What experimental approaches can resolve conflicting data about ZNF503 function across different tissue types?

To resolve conflicting data about ZNF503 function across different tissue types:

• Tissue-specific conditional knockout models:

  • Allows assessment of ZNF503 function in specific tissues without developmental lethality

  • Important since biallelic loss-of-function mutations are hypothesized to be lethal

• Single-cell RNA sequencing:

  • Characterize cell type-specific ZNF503 expression and target genes

  • Particularly useful in heterogeneous tissues like developing eyes or tumors

• Comparative ChIP-seq across tissues:

  • Identify tissue-specific binding patterns of ZNF503

  • Correlate with epigenetic modifications to understand context-dependent function

• Proteomics analysis:

  • Identify tissue-specific interaction partners that may explain diverse functions

  • Analysis of post-translational modifications that might regulate activity

• Isoform-specific analyses:

  • ZNF503 may have multiple protein isoforms (28 kDa, 55 kDa, 63 kDa)

  • The UK 100,000 Genomes Project identified a possible N-terminal truncated shorter isoform starting at aa 79

  • Isoform-specific antibodies or genetic approaches can help distinguish functions

These approaches can help reconcile apparently contradictory roles of ZNF503 across different biological contexts .

How can researchers investigate the relationship between ZNF503 and its antisense RNAs (ZNF503-AS1, ZNF503-AS2) in regulatory networks?

To investigate the relationship between ZNF503 and its antisense RNAs:

• Dual RNA/protein detection:

  • Simultaneous detection of ZNF503 protein (using antibodies) and ZNF503-AS RNAs (using RNA FISH)

  • Correlative analysis of expression patterns in tissues or cell populations

• Antisense RNA manipulation:

  • Overexpression or knockdown of ZNF503-AS1/2 followed by ZNF503 protein analysis

  • ZNF503-AS1 has been shown to regulate retinal pigment epithelium differentiation by downregulating ZNF503 expression

• RNA-protein interaction studies:

  • RNA immunoprecipitation (RIP) to determine if ZNF503-AS RNAs interact with proteins that regulate ZNF503

  • Crosslinking immunoprecipitation (CLIP) to map interaction sites

• Functional genomics:

  • CRISPR-Cas9 editing of antisense RNA loci while preserving ZNF503 coding sequence

  • Analysis of effects on ZNF503 expression and phenotypic outcomes

• Clinical correlation studies:

  • ZNF503-AS1 levels correlate with diabetic retinopathy incidence and can regulate TGF-β1 expression

  • Systematic analysis of ZNF503 and its antisense RNAs across patient cohorts

These approaches can help elucidate the regulatory relationship between ZNF503 and its antisense RNAs in development and disease .

What are common issues when using ZNF503 antibodies and how can they be resolved?

Common issues with ZNF503 antibodies and their solutions include:

Always validate antibodies using positive controls (293T, Jurkat, MCF7, or PC-3 cell lysates) and include appropriate negative controls in your experimental design .

What are the best practices for preserving antibody functionality during storage and handling?

Best practices for preserving ZNF503 antibody functionality:

• Storage recommendations:

  • For immediate use, store at 4°C for up to two weeks

  • For long-term storage, divide into small aliquots (≥20 μL) and freeze at -20°C or -80°C

  • Avoid repeated freeze-thaw cycles that can degrade antibody activity

• Handling guidelines:

  • Centrifuge briefly before opening the vial to collect all liquid

  • Use sterile techniques when handling antibody solutions

  • Store in appropriate buffer conditions (typically with glycerol or carrier proteins)

• Working dilution preparation:

  • Prepare fresh working dilutions on the day of use

  • Return stock solutions to appropriate storage temperatures immediately

  • Use appropriate diluents (typically PBS with 0.1% BSA or similar carrier protein)

• Quality control:

  • Periodically test antibody activity using positive controls

  • Note lot numbers and maintain consistent sourcing when possible

  • The recommended shelf life for some ZNF503 antibodies is 1 year from date of receipt

Following these practices will help maintain antibody specificity and sensitivity throughout your research project .

How can ZNF503 antibodies contribute to understanding developmental disorders and potential therapeutic interventions?

ZNF503 antibodies can contribute to understanding developmental disorders through:

• Developmental timing studies:

  • ZNF503 is dynamically expressed during mouse embryogenesis and eye development

  • Antibodies can track protein expression patterns across developmental stages

• Lineage commitment analysis:

  • ZNF503 regulates RPE differentiation; loss leads to conversion to neural-retina-like lineages

  • Antibodies can monitor differentiation states in developmental models

• Genetic disease correlations:

  • ZNF503 is highly constrained with a pLI score of 0.95, suggesting importance in development

  • Antibodies can verify protein expression in patient-derived cells with ZNF503 variants

• Therapeutic target validation:

  • ZNF503 is involved in multiple disease processes including cancer progression

  • Antibodies can validate target engagement in drug development

• Gene therapy monitoring:

  • For developmental disorders caused by ZNF503 dysfunction

  • Antibodies can track restoration of normal protein levels following intervention

These applications could advance understanding of developmental disorders like uveal coloboma and inform potential therapeutic approaches .

What role might ZNF503 play in cancer biology, and how can researchers design experiments to investigate this?

ZNF503's role in cancer biology can be investigated through:

• Expression correlation with clinical outcomes:

  • ZNF503 upregulation in HCC correlates with advanced TNM stage, venous invasion, and poor survival

  • 5-year survival data shows ZNF503 as a potential prognostic marker

  Clinical Parameter	ZNF503 high (n=48)	ZNF503 low (n=47)	P value
  TNM stage (III+IV)	16	5	0.008
  Venous infiltration	11	3	0.023

• Functional studies of cancer-related processes:

  • ZNF503 promotes migration, invasion, and EMT progression in HCC cells

  • Design gain/loss-of-function experiments in cancer cell lines with antibody validation

• Mechanistic pathway analysis:

  • ZNF503 represses GATA3 by direct promoter binding in HCC cells

  • ZNF503 is regulated by miR-495 in HCC, with inverse correlation in tissues

  • Design ChIP experiments using ZNF503 antibodies to identify cancer-specific targets

• Therapeutic targeting approaches:

  • Design experiments to test if blocking ZNF503 function inhibits cancer progression

  • Use antibodies to monitor ZNF503 levels following experimental treatments

• Clinical translation research:

  • Develop tissue microarray studies using ZNF503 antibodies across cancer types

  • Correlate with treatment response and clinical outcomes

These approaches can help establish ZNF503 as a candidate for diagnostic, prognostic, and therapeutic applications in cancer .

What are potential applications of ZNF503 antibodies in single-cell analysis technologies?

Potential applications of ZNF503 antibodies in single-cell technologies include:

• Mass cytometry (CyTOF):

  • Incorporating metal-conjugated ZNF503 antibodies into CyTOF panels

  • Allows simultaneous detection of ZNF503 with numerous other proteins

  • Useful for analyzing heterogeneous populations in developmental processes or tumors

• Single-cell Western blotting:

  • Analysis of ZNF503 expression variations at the single-cell level

  • Can detect different isoforms or post-translational modifications

• Imaging mass cytometry:

  • Spatial detection of ZNF503 in tissue sections with subcellular resolution

  • Correlation with tissue architecture and microenvironment

• Proximity ligation assays:

  • Detect protein-protein interactions involving ZNF503 in single cells

  • Particularly useful for studying transcriptional complexes

• CODEX multiplexed imaging:

  • Integration of ZNF503 antibodies into highly multiplexed imaging panels

  • Spatial mapping of ZNF503 expression relative to multiple cell markers

These emerging technologies can reveal previously undetectable heterogeneity in ZNF503 expression and function at single-cell resolution .

How might the study of ZNF503 and related zinc finger proteins advance our understanding of transcriptional regulation in development and disease?

The study of ZNF503 and related zinc finger proteins can advance understanding of transcriptional regulation through:

• Comparative functional genomics:

  • ZNF503 belongs to the NET subfamily related to the Sp family of transcription factors

  • Systematic comparison of binding sites and regulatory networks across family members

• Structural biology approaches:

  • Determination of ZNF503 DNA-binding specificity and protein structure

  • Development of structure-based modulators of function

• Synthetic biology applications:

  • Engineering ZNF503-based transcriptional regulators for precise control of gene expression

  • Potential therapeutic applications in diseases with dysregulated gene expression

• Evolutionary developmental biology:

  • ZNF503 is evolutionarily conserved

  • Comparative studies across species can reveal fundamental mechanisms of development

• Systems biology integration:

  • Positioning ZNF503 within complex gene regulatory networks

  • Mathematical modeling of developmental processes influenced by zinc finger proteins

These approaches can provide insights into fundamental principles of transcriptional regulation and potentially identify novel therapeutic targets for developmental disorders and cancer .