zgc:153521 Antibody

What is zgc:153521 and why is it important in zebrafish research?

zgc:153521 is a secreted protein in zebrafish (Danio rerio) with UniProt Number A0JMH2 and Entrez Gene ID 777620 . This protein is part of the zebrafish gene collection, which contains fully sequenced and annotated genes. The importance of zgc:153521 stems from its potential role in developmental processes, as zebrafish serve as an excellent model organism for studying vertebrate development and disease mechanisms. Zebrafish models have been particularly valuable in studying conditions like tuberculosis , where specialized antibodies enable tracking of protein expression patterns during development and disease progression.

What are the physical characteristics and applications of zgc:153521 antibody?

The commercially available zgc:153521 antibody is typically a rabbit polyclonal antibody produced against recombinant Danio rerio (zebrafish) zgc:153521 protein . This antibody is primarily used for:

The antibody is typically purified by antigen affinity chromatography and is supplied with positive controls (recombinant antigen) and negative controls (pre-immune serum) to validate experimental results .

How should researchers optimize zgc:153521 antibody protocols for zebrafish tissue samples?

When using zgc:153521 antibody with zebrafish samples, researchers should:

• Sample preparation: For adult zebrafish studies, consider using longitudinal tissue sections similar to those employed in tuberculosis model studies . For embryonic studies, whole-mount preparations may be more appropriate.

• Fixation optimization: Test both 4% paraformaldehyde (PFA) and alternative fixatives, as epitope accessibility can vary significantly. For developmental studies tracking protein expression over time, consistent fixation protocols are essential.

• Antibody validation: Perform Western blot analysis to confirm specific binding at the expected molecular weight. Include appropriate controls:

  • Positive control: recombinant zgc:153521 protein

  • Negative control: pre-immune serum

  • Tissue-specific negative control: tissues known not to express the target

• Signal detection: For fluorescent applications, minimize autofluorescence by comparing multiple secondary antibody conjugates (Cy3 has been used successfully with similar zebrafish antibodies ).

What are the recommended controls for experiments using zgc:153521 antibody?

Proper experimental controls are critical for ensuring reliable results:

• Technical controls:

  • No primary antibody control

  • Isotype control (irrelevant rabbit IgG at matching concentration)

  • Blocking peptide competition assay (pre-incubating antibody with immunizing peptide)

• Biological controls:

  • Wild-type vs. mutant comparisons

  • Consider using genetic approaches like those employed in il10e46 mutants in zebrafish tuberculosis models

  • Developmental time series to establish expression patterns

  • Tissue-specific positive and negative controls

• Quantification controls: Include standardized proteins of known concentration when performing quantitative Western blot analysis.

How can zgc:153521 antibody be integrated into multi-parameter analyses?

For comprehensive studies of developmental processes or disease mechanisms:

• Dual immunofluorescence: zgc:153521 antibody can be paired with other antibodies (e.g., markers of inflammation or cell death) to study protein co-localization, following protocols similar to those used in zebrafish tuberculosis models .

• Transcriptome correlation: Combine antibody-based protein detection with RNA-seq approaches, as demonstrated in zebrafish infection models where whole genome-level transcriptome analysis revealed relationships between protein expression and gene regulation .

• Cross-platform validation: Validate antibody-based findings with complementary approaches:

  • qRT-PCR for gene expression

  • In situ hybridization for spatial localization

  • Transgenic reporter lines for real-time visualization

What strategies can address common challenges with zgc:153521 antibody experiments?

• High background signal:

  • Increase blocking concentration (5-10% normal serum)

  • Optimize antibody concentration through titration experiments

  • Try alternative blocking reagents (BSA, milk, commercial blockers)

  • Increase washing duration and frequency

• Weak or no signal:

  • Test multiple antigen retrieval methods (heat-induced vs. enzymatic)

  • Modify antibody incubation conditions (time, temperature, concentration)

  • Ensure protein is not lost during sample preparation

  • Verify that the target epitope is accessible under your experimental conditions

• Cross-reactivity issues:

  • Perform peptide competition assays

  • Test antibody on tissues known to lack the target

  • Consider pre-absorption with related proteins

How can zgc:153521 antibody contribute to zebrafish disease model development?

Zebrafish are valuable model organisms for studying human diseases, particularly infectious diseases like tuberculosis and developmental disorders. The zgc:153521 antibody can be employed to:

• Track protein expression changes during disease progression, as demonstrated in M. marinum infection studies in zebrafish where tissue sections were analyzed at different timepoints .

• Evaluate treatment efficacy by monitoring protein expression changes following therapeutic interventions.

• Study protein localization in response to stressors, similar to studies examining granuloma formation in zebrafish tuberculosis models .

• Investigate signal pathway activation, particularly in immune response studies where antibody detection can complement transcriptional analysis of immune response elements .

What considerations are important when using zgc:153521 antibody in developmental studies?

When studying developmental processes:

• Temporal resolution: Plan collection timepoints carefully to capture dynamic expression changes. In zebrafish embryos, consider that zygotic transcription begins around 3 hours post-fertilization (hpf) .

• Spatial distribution: Whole-mount preparations can provide valuable spatial information, but may require extended antibody incubation times and specialized permeabilization protocols.

• Quantification challenges: Develop consistent imaging and quantification strategies, potentially using automated image analysis tools to reduce bias.

• Developmental context: Interpret findings in light of concurrent developmental events, as protein function may vary depending on developmental stage.

How should researchers quantify and analyze zgc:153521 expression data?

Quantitative analysis of zgc:153521 expression requires:

• Western blot quantification:

  • Use appropriate loading controls (housekeeping proteins)

  • Employ standard curves with recombinant protein for absolute quantification

  • Apply digital image analysis with software that accounts for non-linear signal response

• Immunohistochemistry quantification:

  • Establish consistent image acquisition parameters

  • Develop tissue-specific scoring systems (e.g., H-score, Allred score)

  • Consider automated analysis tools to reduce bias

• Statistical approaches:

  • Account for biological and technical variability

  • Apply appropriate statistical tests based on data distribution

  • Consider sample size calculations to ensure adequate power

What bioinformatic resources can complement zgc:153521 antibody studies?

Multiple bioinformatic resources can enhance interpretation of antibody-based findings:

• Sequence and structure databases:

  • UniProt (A0JMH2) for protein sequence information

  • NCBI resources for gene information (Entrez Gene ID: 777620)

  • The Patent and Literature Antibody Database (PLAbDab) for related antibody information

• Functional prediction tools:

  • Gene Ontology resources for predicted functions

  • Pathway analysis tools to place findings in broader biological context

  • Comparative genomics to identify homologs in other species

• Integration with transcriptomic data:

  • RNA-seq datasets from zebrafish developmental stages or disease models

  • Single-cell sequencing data for cell-type specific expression patterns

  • Public repositories like GEO for comparative analysis