ZNF215 Antibody

What is ZNF215 and what are its known biological functions?

ZNF215 is a zinc finger protein encoded by the ZNF215 gene (NCBI Gene ID: 7762). It is also known by several synonyms including BAZ2, ZKSCAN11, ZSCAN43, and BAZ-2 . As a zinc finger protein, ZNF215 likely functions as a transcription factor involved in gene regulation. ZNF215 has approximately 3,062 functional associations with various biological entities spanning 8 categories, including molecular profiles, organisms, chemicals, functional terms, diseases, phenotypes, structural features, and cell types, suggesting its involvement in multiple biological processes .

The protein has a calculated molecular weight of approximately 60 kDa based on its 517 amino acid sequence, though it is typically observed at 55-60 kDa in experimental conditions .

What applications are ZNF215 antibodies validated for?

ZNF215 antibodies have been validated for several research applications:

ZNF215 antibodies have shown reactivity with human and mouse samples, with specific detection confirmed in mouse brain tissue for WB and HepG2 cells for IF/ICC applications .

What is the difference between monoclonal and polyclonal ZNF215 antibodies?

The search results indicate availability of both monoclonal and polyclonal ZNF215 antibodies, each with distinct characteristics:

Polyclonal ZNF215 antibodies:

• Typically raised in rabbit hosts (e.g., 25502-1-AP)

• Recognize multiple epitopes on the ZNF215 protein

• Offer robust detection across applications like WB and IF/ICC

• Generated using ZNF215 fusion protein immunogens (e.g., Ag22026)

• Particularly useful for detecting native proteins due to recognition of multiple epitopes

Monoclonal ZNF215 antibodies:

• Available in specific clones (e.g., 1E4, 2C11)

• Host organism is typically mouse

• Target a single epitope on the ZNF215 protein

• Provide highly consistent lot-to-lot reproducibility

• Particularly valuable in antibody pair applications where specificity is critical

The choice between monoclonal and polyclonal depends on the specific research application. For antibody pairs used in ELISA and other quantitative assays, using a combination (e.g., mouse monoclonal capture antibody with rabbit polyclonal detection antibody) provides optimal specificity and sensitivity .

What are the optimal storage conditions for ZNF215 antibodies?

For maintaining optimal activity of ZNF215 antibodies, the following storage conditions are recommended:

• Store at -20°C for long-term storage

• Antibodies remain stable for one year after shipment when properly stored

• Aliquoting is recommended to avoid repeated freeze-thaw cycles, which can degrade antibody quality

• For smaller quantities (20μL sizes), some formulations contain 0.1% BSA as a stabilizer

• Typical storage buffer consists of PBS with 0.02% sodium azide and 50% glycerol at pH 7.3

• Antibodies should be returned to -20°C storage immediately after use

How should I optimize Western blot protocols specifically for ZNF215 detection?

When performing Western blot for ZNF215 detection, consider these optimization strategies:

• Sample Selection: Mouse brain tissue has been validated as a positive control for ZNF215 Western blot detection .

• Dilution Range: Begin with the recommended dilution range of 1:200-1:1000 . Optimal dilution should be determined empirically for each experimental system.

• Expected Band Size: Look for bands in the 55-60 kDa range, which corresponds to the observed molecular weight of ZNF215 .

• Antibody Selection: For standard WB, a single anti-ZNF215 antibody (such as the rabbit polyclonal 25502-1-AP) can be used . For more sensitive detection, consider using an IP-WB antibody pair system which includes a rabbit polyclonal for immunoprecipitation and a mouse polyclonal for detection .

• Protocol Specificity: For optimal results, follow antibody-specific protocols provided by manufacturers. Specific WB protocols are available for individual antibodies (e.g., ZNF215 antibody 25502-1-AP) .

• System-Specific Optimization: As noted in the product literature, "it is recommended that this reagent should be titrated in each testing system to obtain optimal results" .

What considerations are important when using ZNF215 antibodies for immunofluorescence?

When conducting immunofluorescence (IF) experiments with ZNF215 antibodies:

• Cell Line Selection: HepG2 cells have been validated as a positive control for ZNF215 immunofluorescence detection .

• Dilution Range: Start with the recommended dilution range of 1:10-1:100 for IF/ICC applications .

• Fixation Considerations: Follow specific IF protocols provided by the manufacturer for optimal fixation and permeabilization parameters .

• Protocol Optimization: Given the relatively concentrated antibody usage in IF applications, careful optimization of each step is crucial for specific signal detection while minimizing background.

• Visualization Method: ZNF215 antibodies are typically unconjugated, requiring appropriate secondary antibodies for visualization .

How can I effectively use ZNF215 antibody pairs for protein quantification?

ZNF215 antibody pairs are specifically designed for applications requiring high specificity and sensitivity, such as ELISA and immunoprecipitation followed by Western blot (IP-WB):

For ELISA applications:

• Pair Components: Use matched antibody pairs consisting of:

  • Capture antibody: mouse monoclonal anti-ZNF215 (100 μg)

  • Detection antibody: rabbit purified polyclonal anti-ZNF215 (50 μg)

• Assay Scale: The reagents provided in typical antibody pair sets are sufficient for at least 3-5 x 96 well plates using recommended protocols .

For IP-WB applications:

• Pair Components: Use matched antibody pairs consisting of:

  • IP antibody: rabbit polyclonal anti-ZNF215 (300 μL)

  • WB detection antibody: mouse purified polyclonal anti-ZNF215 (50 μg)

• Methodology: The paired approach allows for initial enrichment of the target protein by immunoprecipitation, followed by specific detection via Western blot, providing enhanced sensitivity compared to direct Western blot.

• Controls: Include appropriate negative controls (isotype-matched non-specific antibodies) to validate specificity of the immunoprecipitation.

What approaches should I consider for studying ZNF215 expression in brain tissues?

Based on the search results, ZNF215 expression has been analyzed in various brain tissues through multiple datasets:

• Available Expression Datasets:

  • Allen Brain Atlas Adult Human Brain Tissue Gene Expression Profiles

  • Allen Brain Atlas Developing Human Brain Tissue Gene Expression Profiles by Microarray

  • Allen Brain Atlas Developing Human Brain Tissue Gene Expression Profiles by RNA-seq

  • Allen Brain Atlas Prenatal Human Brain Tissue Gene Expression Profiles

• Experimental Approaches:

  • Histological Analysis: Immunohistochemistry using validated anti-ZNF215 antibodies can reveal spatial distribution within brain regions.

  • Expression Correlation: Compare ZNF215 protein expression (via Western blot or IHC) with gene expression data from transcriptomic databases to understand regulation mechanisms.

  • Developmental Studies: Consider temporal expression patterns using prenatal and developing brain datasets to investigate ZNF215's role in neurodevelopment.

• Technical Considerations:

  • For Western blot analysis, mouse brain tissue has been validated for ZNF215 detection .

  • Consider using standardized values from existing expression datasets to identify regions of interest before designing targeted protein studies.

How do I troubleshoot non-specific binding when using ZNF215 antibodies?

When encountering non-specific binding with ZNF215 antibodies, consider these troubleshooting approaches:

• Antibody Selection:

  • For applications requiring highest specificity, consider using monoclonal antibodies (e.g., clones 1E4 or 2C11) .

  • For certain applications, antibody pairs may provide better specificity than single antibodies .

• Blocking Optimization:

  • Ensure adequate blocking steps in your protocol.

  • Optimize blocking buffer composition based on the specific sample type.

• Dilution Optimization:

  • Test a range of antibody dilutions, starting with manufacturer recommendations (e.g., 1:200-1:1000 for WB) .

  • Determine optimal dilution empirically for each experimental system, as explicitly recommended by manufacturers .

• Sample-Dependent Considerations:

  • As noted in the product information, results can be sample-dependent .

  • Check validation data galleries provided by manufacturers for sample-specific recommendations.

• Protocol Adherence:

  • Follow antibody-specific protocols provided by manufacturers .

  • Pay particular attention to washing steps to reduce background signal.

What controls should I include in ZNF215 antibody experiments?

Proper experimental controls are essential for valid interpretation of ZNF215 antibody results:

• Positive Controls:

  • For Western blot: Mouse brain tissue has been validated for ZNF215 detection .

  • For IF/ICC: HepG2 cells have been validated for ZNF215 detection .

• Negative Controls:

  • Primary antibody omission: Include samples processed identically but without primary ZNF215 antibody.

  • Isotype controls: Use non-specific antibodies of the same isotype (e.g., Rabbit IgG for polyclonal rabbit anti-ZNF215).

• Validation Controls:

  • Consider using siRNA or CRISPR knockdown of ZNF215 to confirm antibody specificity.

  • For antibody pairs, include single antibody controls to evaluate non-specific binding.

• Technical Controls:

  • Include loading controls for Western blot (e.g., housekeeping proteins).

  • For IF/ICC, include nuclear counterstains to assess cellular morphology.

How should I design experiments to study ZNF215 protein-protein interactions?

To investigate ZNF215 protein interactions:

• Immunoprecipitation Approach:

  • Utilize specialized IP-WB antibody pairs designed for ZNF215 .

  • The rabbit polyclonal component (300 μL) can effectively capture ZNF215 and its interacting partners.

  • The mouse polyclonal component (50 μg) can then detect ZNF215 in the precipitated complex via Western blot.

• Co-immunoprecipitation:

  • After immunoprecipitating ZNF215, probe for suspected interaction partners using specific antibodies.

  • Consider using crosslinking approaches to stabilize transient interactions.

• Reverse Approach:

  • Immunoprecipitate suspected interaction partners and probe for ZNF215.

  • Compare results from both directions to validate interactions.

• Functional Association Context:

  • Consider the 3,062 functional associations with biological entities spanning 8 categories that have been identified for ZNF215 .

  • Design experiments targeting proteins within relevant functional networks.

What considerations are important when selecting between different ZNF215 antibody formats?

When selecting ZNF215 antibodies for specific applications:

• Application-Specific Selection:

  Application	Recommended Format	Justification
  Western Blot	Rabbit polyclonal or IP-WB pair	Validated in mouse brain tissue, provides specific detection at 55-60 kDa
  IF/ICC	Rabbit polyclonal	Validated in HepG2 cells with established dilution range
  ELISA	Matched antibody pair	Capture/detection system optimized for quantification
  IP	Rabbit polyclonal	Higher affinity for native protein conformation

• Host Organism Considerations:

  • Rabbit-hosted antibodies show reactivity with both human and mouse ZNF215 .

  • Consider the host when designing multi-antibody experiments to avoid cross-reactivity.

• Clonality Decision Factors:

  • For consistent lot-to-lot reproducibility, consider monoclonal antibodies (e.g., clones 1E4, 2C11) .

  • For detection of native proteins across species, polyclonal antibodies may offer broader epitope recognition.

• Format-Specific Advantages:

  • Standalone antibodies: Simplicity of use in standard applications.

  • Antibody pairs: Enhanced specificity and sensitivity in complex samples.

  • IP-WB pairs: Specialized for enrichment and detection of low-abundance targets.

How can I verify the specificity of my ZNF215 antibody results?

Verifying antibody specificity is crucial for reliable ZNF215 research:

• Band Pattern Analysis:

  • Confirm that the observed molecular weight matches the expected 55-60 kDa range for ZNF215 .

  • The calculated molecular weight based on the 517 amino acid sequence is approximately 60 kDa .

• Positive Control Verification:

  • Compare results with validated positive controls (mouse brain tissue for WB, HepG2 cells for IF/ICC) .

  • Consistent results across these controls support antibody specificity.

• Genetic Approaches:

  • Consider RNAi verification, as some ZNF215 antibodies are validated for RNAi applications .

  • CRISPR/Cas9 knockout can provide definitive validation of antibody specificity.

• Multi-Antibody Validation:

  • Compare results using different antibodies targeting distinct epitopes of ZNF215.

  • Concordant results across multiple antibodies strongly support specificity.

• Specificity Testing:

  • For immunogenic peptide-raised antibodies, consider peptide competition assays.

  • Preincubation of the antibody with excess target peptide should abolish specific binding.

How do I correlate ZNF215 protein expression with gene expression data?

To meaningfully correlate ZNF215 protein and gene expression:

• Available Expression Resources:

  • Utilize gene expression data from:

    • Allen Brain Atlas resources (Adult, Developing, and Prenatal Human Brain Tissue)

    • BioGPS Cell Line Gene Expression Profiles

    • BioGPS Human Cell Type and Tissue Gene Expression Profiles

• Methodology Approach:

  • Quantify ZNF215 protein levels using validated antibodies in Western blot (densitometry) or ELISA.

  • Obtain corresponding mRNA expression data from appropriate databases.

  • Perform statistical correlation analysis between protein and mRNA levels.

• Interpreting Discrepancies:

  • Consider post-transcriptional and post-translational regulation mechanisms.

  • Protein stability differences can lead to poor correlation with mRNA levels.

  • Time-course studies may reveal delays between mRNA expression and protein accumulation.

• Cell Type Considerations:

  • Account for potential cell type-specific expression patterns.

  • Single-cell approaches may reveal heterogeneity masked in bulk tissue analysis.