pop1 Antibody

What are the primary applications for POP1 antibody in research?

POP1 antibody is versatile for multiple experimental applications, with validated performance in Western Blot (WB), Immunoprecipitation (IP), Immunohistochemistry (IHC), Immunofluorescence (IF/ICC), RNA Immunoprecipitation (RIP), and ELISA assays. Published literature confirms its effectiveness across these applications, with particularly robust performance in Western Blot applications where it has been cited in at least seven publications . The antibody's versatility makes it valuable for investigating POP1's role in various cellular processes, particularly in ribosome biogenesis and RNA processing pathways .

Which species does POP1 antibody reliably detect?

POP1 antibody (12029-1-AP) has demonstrated consistent reactivity with human, mouse, and rat samples across multiple experimental contexts . While tested reactivity confirms these three species, cited reactivity in published literature primarily focuses on human samples . For researchers working with other mammalian models, preliminary validation is recommended before proceeding with large-scale experiments.

What is the recommended dilution range for POP1 antibody across different applications?

Different applications require specific dilution ranges for optimal results as detailed in the following table:

It is essential to note that optimal dilution is sample-dependent, and researchers should perform preliminary dilution series experiments to determine the ideal concentration for their specific experimental conditions .

What is the molecular weight of POP1 protein that should be detected?

When working with POP1 antibody, researchers should expect to observe bands at approximately 115 kDa, which corresponds to the observed molecular weight of the POP1 protein . The calculated molecular weight based on amino acid composition is 115 kDa (1024 amino acids), which aligns with the observed molecular weight in experimental conditions . This consistency between calculated and observed weights provides confidence in antibody specificity.

How can POP1 antibody be used to investigate the role of POP1 in cancer research?

Recent research has revealed significant implications for POP1 in cancer biology, particularly in breast cancer (BC). Studies have demonstrated that POP1 expression is upregulated in BC tissue compared to normal breast tissue, with a higher mutation rate . This makes POP1 antibody a valuable tool for cancer researchers to:

• Evaluate POP1 expression levels in tumor samples using IHC (recommended dilution 1:400-1:1600)

• Investigate POP1's association with poor prognosis through protein expression analysis

• Explore POP1's role in immune microenvironment regulation using IP and co-IP experiments

• Assess the correlation between POP1 expression and response to immunotherapy

Notably, patients with high POP1 expression were found more likely to respond to immunotherapy, suggesting POP1's potential role as a biomarker for predicting immunotherapy response . Western blot analysis using POP1 antibody can help stratify patient samples based on expression levels to investigate this correlation further.

What methodological approaches can address inconsistent results when using POP1 antibody in IHC applications?

When inconsistent IHC results occur with POP1 antibody, consider the following methodological refinements:

How can researchers validate POP1 antibody specificity in their experimental systems?

Validating antibody specificity is crucial for result interpretation. For POP1 antibody, implement the following validation strategy:

• Positive and negative control samples: The antibody has demonstrated positive Western blot detection in HeLa and HepG2 cells, making these appropriate positive controls . Include cell lines known not to express POP1 as negative controls.

• Knockdown/knockout validation: Published applications confirm the antibody's effectiveness in KD/KO experimental designs . Generate POP1 knockdown/knockout samples to verify signal disappearance.

• Multiple antibody approach: Consider validating results with another POP1 antibody targeting a different epitope, such as comparing results between polyclonal (CAB5961) and monoclonal antibodies .

• Peptide competition assay: Pre-incubate the antibody with purified POP1 protein or immunogenic peptide before application to verify signal reduction.

• Expected molecular weight confirmation: Verify that detected bands appear at the expected 115 kDa position .

What evidence supports POP1 as a diagnostic biomarker, and how can researchers evaluate its potential?

Meta-analysis of multiple cohorts has demonstrated POP1's significant diagnostic potential in breast cancer. The diagnostic accuracy assessed through ROC curve analysis revealed impressive results across different cohorts:

The combined meta-analysis demonstrated robust diagnostic value with a combined AUC of 0.90 (95% CI: 0.87-0.92) . Researchers evaluating POP1 as a diagnostic biomarker should:

• Apply POP1 IHC staining to tissue microarrays containing both tumor and adjacent normal tissue

• Correlate POP1 expression with established diagnostic markers

• Implement quantitative image analysis to establish standardized scoring systems

• Validate findings across multiple independent cohorts to minimize bias

The high stability of meta-analysis results when excluding any individual cohort indicates robust diagnostic potential that warrants further investigation .

How does POP1 influence immune infiltration, and how can this be studied using POP1 antibody?

Research indicates that POP1 plays a significant role in immune infiltration patterns within the tumor microenvironment. To investigate this relationship using POP1 antibody:

• Multiplex immunofluorescence: Combine POP1 antibody (1:50-1:500 dilution range for IF) with markers for specific immune cell populations to visualize co-localization patterns .

• Correlation analysis: Following IHC or IF staining with POP1 antibody, perform quantitative analysis correlating POP1 expression levels with immune cell densities.

• Flow cytometry: Use flow cytometry to study associations between POP1 expression and immune cell phenotypes in dissociated tumor samples.

• Co-immunoprecipitation: Employ POP1 antibody for IP (using 0.5-4.0 μg for 1.0-3.0 mg of total protein) to identify potential interaction partners within immune signaling pathways .

Studies utilizing CIBERSORT analysis have revealed significant correlations between POP1 expression and specific immune cell infiltration patterns, with differential infiltration observed between high and low POP1 expression groups . Researchers should consider both direct approaches using antibody-based detection and computational methods to comprehensively characterize these relationships.

What storage and handling precautions are critical for maintaining POP1 antibody activity?

To preserve antibody performance over time, researchers should adhere to these evidence-based handling protocols:

• Storage temperature: Store POP1 antibody at -20°C where it remains stable for one year after shipment .

• Buffer composition: The antibody is supplied in PBS with 0.02% sodium azide and 50% glycerol at pH 7.3, which helps maintain stability during storage .

• Aliquoting considerations: For the standard storage temperature (-20°C), aliquoting is generally unnecessary, which simplifies handling protocols .

• Freeze-thaw cycles: Despite not requiring aliquoting, minimizing freeze-thaw cycles is recommended to preserve epitope recognition capability.

• Working solution preparation: For working solutions, dilute using fresh, sterile buffer solutions and use within 24 hours for optimal performance.

• Contamination prevention: Use sterile technique when handling antibody solutions to prevent microbial contamination that could degrade antibody performance.

How can researchers optimize Western blot protocols specifically for POP1 detection?

For optimal Western blot results with POP1 antibody, implement these methodological refinements:

• Sample preparation: Given POP1's high molecular weight (115 kDa), use lower percentage gels (8-10%) for better resolution and transfer efficiency.

• Protein loading: Load sufficient protein (20-50 μg total protein per lane) to ensure detection of less abundant POP1.

• Transfer conditions: Implement extended transfer times or reduced voltage for high molecular weight proteins to ensure complete transfer.

• Blocking optimization: Test different blocking agents (5% non-fat milk versus BSA) to determine optimal signal-to-noise ratio.

• Antibody concentration: While the recommended dilution range is 1:2000-1:16000, begin with 1:5000 and adjust based on signal intensity .

• Validated cell lines: HeLa and HepG2 cells have been confirmed to express detectable levels of POP1 and serve as excellent positive controls .

• Detection system selection: For low abundance targets, consider using enhanced chemiluminescence (ECL) systems with extended exposure times or fluorescence-based detection for quantitative analysis.

What considerations should researchers make when designing immunoprecipitation experiments with POP1 antibody?

For successful immunoprecipitation of POP1 and its interaction partners:

• Antibody amount optimization: The recommended range is 0.5-4.0 μg antibody for 1.0-3.0 mg of total protein lysate, but this should be empirically determined for each experimental system .

• Lysis buffer selection: Choose a lysis buffer that preserves protein-protein interactions while effectively solubilizing membrane components (e.g., RIPA buffer with reduced SDS concentration).

• Pre-clearing step: Implement a pre-clearing step with protein A/G beads to reduce non-specific binding.

• Incubation conditions: Optimize antibody-lysate incubation time and temperature (typically 4°C overnight) to maximize specific binding while minimizing non-specific interactions.

• Washing stringency: Balance between sufficient washing to remove non-specific binding and preserving specific interactions.

• Elution methods: Compare different elution methods (e.g., low pH, high salt, SDS) to identify the most efficient approach for POP1 complexes.

• Negative controls: Include an isotype control (Rabbit IgG) processed identically to evaluate non-specific binding.

HeLa cells have been validated for successful IP experiments with POP1 antibody and can serve as a positive control system when establishing protocols .

How can POP1 antibody contribute to understanding cancer progression mechanisms?

POP1 antibody serves as an essential tool for investigating cancer progression through multiple approaches:

• Proliferation and migration analysis: Research has shown that POP1 promotes proliferation and migration while inhibiting apoptosis in breast cancer cells . Researchers can use POP1 antibody to:

  • Quantify POP1 expression changes during disease progression via IHC or Western blot

  • Correlate expression levels with proliferation markers

  • Evaluate POP1 localization changes during epithelial-mesenchymal transition

• Pathway analysis: POP1 has been significantly correlated with multiple pathways in breast cancer . Using techniques like IF and IP with POP1 antibody enables:

  • Identification of co-localization with pathway components

  • Characterization of POP1-containing protein complexes

  • Evaluation of pathway activation states in relation to POP1 expression

• Mutation impact assessment: With higher mutation rates observed in breast cancer tissue , researchers can:

  • Compare wild-type versus mutant POP1 expression patterns

  • Investigate how mutations affect subcellular localization

  • Assess functional consequences of mutations on POP1's role in RNA processing

What approaches can researchers use to investigate POP1's role in immunotherapy response?

Given that high-POP1 expression patients were more prone to respond to immunotherapy , the following experimental approaches using POP1 antibody are recommended:

• Patient stratification: Use POP1 IHC (1:400-1:1600 dilution) to classify patient samples into high and low expression groups .

• Predictive biomarker validation: Correlate POP1 expression levels with documented immunotherapy response data.

• Mechanism investigation: Combine POP1 antibody with other immune checkpoint markers in multiplex IHC to understand mechanistic relationships.

• In vitro modeling: Develop cell line models with varying POP1 expression levels to investigate direct effects on immune cell activation and function.

• Signaling pathway analysis: Use POP1 antibody in Western blot and IP experiments to characterize signaling pathway alterations that might explain differential immunotherapy response.

• Combination therapy evaluation: Assess how POP1 expression might influence response to combination immunotherapy approaches through preclinical models.

These approaches can help establish whether POP1 expression is merely correlative or mechanistically linked to immunotherapy response, potentially leading to more personalized treatment strategies.

What emerging research areas might benefit from POP1 antibody applications?

Several frontier research areas present opportunities for novel POP1 antibody applications:

• Single-cell analysis: Adapting POP1 antibody for single-cell Western blot or mass cytometry could reveal heterogeneity in POP1 expression within tumor populations.

• Liquid biopsy development: Exploring whether POP1 protein can be detected in circulation using highly sensitive immunoassays might enable non-invasive monitoring.

• Therapeutic targeting: As research reveals POP1's role in cancer progression, antibody-based therapeutic approaches targeting POP1 might become feasible.

• Structural biology: Using POP1 antibody for epitope mapping could contribute to understanding functional domains critical for POP1's role in RNA processing.

• Relationship to RNA biology: Given POP1's function in RNA processing, investigating connections between POP1 dysregulation and RNA metabolism disorders represents an untapped research avenue.