BCL7B Antibody

What is BCL7B and why is it significant in cancer research?

BCL7B (B-Cell CLL/lymphoma 7B) is a protein that has demonstrated significant correlations with various cancer types and immune infiltration processes. Recent pan-cancer analysis has revealed that the BCL7B gene has high diagnostic and prognostic value across multiple cancer types and is related to the infiltration of 24 immune cell subsets in various cancers . BCL7B expression has been specifically observed to correlate with immune cell infiltration in bladder cancer (BLCA), low-grade glioma (LGG), prostate adenocarcinoma (PRAD), and thyroid carcinoma (THCA) among other cancer types. The significance of BCL7B lies in its associations with immunoregulatory pathways, cytokine-cytokine receptor interactions, and neutrophil degranulation processes, suggesting its potential role in tumor microenvironment modulation and cancer progression mechanisms .

What applications are BCL7B antibodies typically used for in research?

BCL7B antibodies are primarily utilized in several key laboratory techniques:

• Western Blotting (WB): For detecting and quantifying BCL7B protein expression in cell or tissue lysates with recommended dilutions ranging from 1:500-2000 .

• Enzyme-Linked Immunosorbent Assay (ELISA): For quantitative detection of BCL7B in solution with recommended dilutions of 1:5000-20000 .

• Immunohistochemistry (IHC): For detecting BCL7B protein in tissue sections with recommended dilutions of 1:50-1:200 .

• Immunofluorescence (IF): For cellular localization studies of BCL7B protein in cells or tissues .

These applications enable researchers to study BCL7B expression patterns, localization, and potential functional implications in various experimental contexts and disease models .

What are the key considerations when selecting a BCL7B antibody for experimental use?

When selecting a BCL7B antibody for research applications, several critical factors should be considered to ensure experimental success:

• Species Reactivity: Verify that the antibody reacts with your species of interest. Available BCL7B antibodies demonstrate reactivity to human and mouse samples, with some also showing cross-reactivity with rat samples .

• Clonality: Choose between polyclonal antibodies (which recognize multiple epitopes) or monoclonal antibodies (which recognize a single epitope) based on your specific application needs. Polyclonal antibodies may provide higher sensitivity while monoclonals offer greater specificity .

• Host Species: Consider the host species (typically rabbit or mouse) particularly when designing multi-color experiments to avoid secondary antibody cross-reactivity issues .

• Validated Applications: Confirm that the antibody has been validated for your specific application (WB, ELISA, IHC, or IF) with supporting validation data .

• Epitope/Immunogen Information: Understanding the specific region of BCL7B that the antibody targets can be important for certain applications - some antibodies target specific amino acid regions (e.g., AA 124-202, AA 137-164) .

• Storage and Handling Requirements: Most BCL7B antibodies require storage at -20°C for long-term preservation, with short-term storage at 4°C for up to one month, avoiding repeated freeze-thaw cycles .

What are the recommended storage and handling procedures for BCL7B antibodies?

Proper storage and handling of BCL7B antibodies are essential for maintaining their functionality and experimental reliability:

• Long-term Storage: Store at -20°C for optimal stability and antibody performance for up to one year .

• Short-term Storage: For frequent use, storage at 4°C for up to one month is acceptable but should be limited to minimize degradation .

• Freeze-thaw Cycles: Avoid repeated freeze-thaw cycles which can degrade antibody quality and affect binding efficiency. Aliquoting antibodies upon receipt can mitigate this issue .

• Storage Buffer Composition: BCL7B antibodies are typically stored in PBS containing 50% glycerol and 0.02% sodium azide, which helps maintain stability .

• Working Dilution Preparation: When preparing working dilutions, use fresh buffer solutions and consider adding protein stabilizers for dilutions that will be stored for more than a day.

• Handling Safety: Exercise appropriate precautions when handling antibodies containing sodium azide, as it is toxic and can form explosive compounds in metal plumbing systems .

Following these guidelines will ensure optimal antibody performance and reproducibility across experiments.

How does BCL7B expression correlate with immune cell infiltration across different cancer types?

BCL7B expression demonstrates significant correlations with immune cell infiltration across multiple cancer types, with particularly notable patterns:

*Specific p-values not provided in the source material for PRAD and THCA

Beyond these four cancer types that show correlations with all six immune cell types, BCL7B expression has been found to correlate with varying degrees of immune cell infiltration across 39 different tumor types including ACC, BRCA, CESC, COAD, DLBC, ESCA, GBM, HNSC, KICH, KIRC, LAML, LIHC, LUAD, LUSC, MESO, OV, PAAD, PCPG, READ, SARC, SKCM, STAD, TGCT, THYM, UCEC, and UVM .

When investigating BCL7B's relationship with tumor microenvironment, researchers should consider these cancer-specific patterns of correlation and design experiments accordingly to validate these bioinformatic findings using BCL7B antibodies in relevant cell or tissue types.

What methodological approaches can resolve contradictory data when studying BCL7B using different antibody clones?

When confronted with contradictory results using different BCL7B antibody clones, researchers should implement a systematic validation approach:

• Epitope Mapping Comparison:

  • Compare the epitope regions targeted by different antibody clones. Differences in BCL7B detection may occur because antibodies recognize different domains (e.g., some target AA 124-202 while others target AA 137-164 or other regions) .

  • Design experiments using antibodies targeting distinct epitopes to obtain complementary data.

• Multi-technique Validation:

  • Employ orthogonal techniques (WB, ELISA, IHC, IF) with the same sample to validate findings.

  • For each technique, optimize antibody concentration using dilution series (e.g., 1:500-2000 for WB; 1:5000-20000 for ELISA; 1:50-200 for IHC) .

• Control Implementation:

  • Include positive controls from tissues known to express BCL7B and negative controls where expression is absent.

  • Perform knockdown/knockout experiments to verify antibody specificity.

  • Use blocking peptides specific to the antibody's immunogen to confirm binding specificity .

• Clone Performance Comparison:

  • Directly compare monoclonal and polyclonal antibodies against the same samples.

  • Monoclonal antibodies (e.g., 6D2, 4G1 clones) may provide greater specificity, while polyclonals may offer broader epitope recognition .

• Cross-species Comparative Analysis:

  • Test antibody performance across species (human, mouse, rat) when contradictions arise to identify species-specific variations .

How should researchers interpret BCL7B antibody data in the context of tumor immune microenvironment studies?

Interpretation of BCL7B antibody data in tumor immune microenvironment studies requires consideration of several integrated factors:

What experimental controls are essential when using BCL7B antibodies in cancer research applications?

Implementing rigorous controls is critical when using BCL7B antibodies in cancer research to ensure data validity and reproducibility:

• Specificity Controls:

  • Positive Tissue Controls: Include tissues with known BCL7B expression (based on published data) to confirm detection capability.

  • Negative Tissue Controls: Include tissues with minimal/no BCL7B expression to assess background and non-specific binding.

  • Blocking Peptide Controls: Use the immunogenic peptide (available for purchase from manufacturers) to pre-absorb the antibody and verify binding specificity .

  • Genetic Controls: When possible, include BCL7B knockdown/knockout samples as the ultimate specificity control.

• Technical Controls:

  • Isotype Controls: Include matched isotype IgG (typically rabbit IgG for polyclonal antibodies) to assess non-specific binding .

  • Secondary Antibody-Only Controls: Omit primary antibody to identify potential non-specific binding of detection systems.

  • Dilution Series: Test multiple antibody dilutions to optimize signal-to-noise ratio (recommended: WB 1:500-2000; ELISA 1:5000-20000; IHC 1:50-200) .

• Cross-validation Controls:

  • Multiple Antibody Clones: When possible, validate key findings using different antibody clones targeting distinct epitopes .

  • Orthogonal Methods: Confirm protein expression using complementary techniques (e.g., if using IHC, confirm with WB) .

• Biological Context Controls:

  • Normal/Tumor Paired Samples: Compare BCL7B expression between matched normal and tumor tissues from the same patient when possible.

  • Cancer Subtype Comparisons: Include multiple cancer subtypes known to have differential BCL7B expression.

  • Immune Infiltration Correlation: Given BCL7B's relationship with immune infiltration, consider co-staining with immune cell markers to establish biological context .

• Experimental Condition Controls:

  • Technical Replicates: Include replicate samples to assess experimental variability.

  • Fixed Analysis Parameters: Maintain consistent imaging parameters, exposure times, and quantification methods across all samples.

  • Treatment Time Course: When studying effects of interventions on BCL7B expression, include appropriate time-point controls.

Implementing these comprehensive controls will significantly strengthen the reliability of BCL7B antibody-derived data in cancer research applications.

How can BCL7B antibodies be effectively used to investigate its role in cancer immune evasion mechanisms?

To effectively investigate BCL7B's role in cancer immune evasion using antibody-based approaches, researchers should implement a multi-level experimental strategy:

• Expression Profiling in Immune-Resistant Models:

  • Compare BCL7B expression between immunotherapy-responsive and resistant tumors using validated antibody dilutions (IHC 1:50-200) .

  • Correlate BCL7B levels with patient response to immunotherapy in clinical samples.

  • Utilize both tissue microarrays and whole-section IHC to address heterogeneity concerns.

• Immune Checkpoint Correlation Studies:

  • Design multi-color immunofluorescence panels combining BCL7B antibodies with antibodies against relevant immune checkpoints.

  • Based on bioinformatic data showing BCL7B correlation with 47 immune checkpoints across 39 cancer types, prioritize checkpoints with strongest correlations for co-expression studies .

  • Implement dual-IHC or sequential immunofluorescence to visualize co-localization patterns.

• Immune Cell Infiltration Assessment:

  • Design multiplexed immunofluorescence panels to simultaneously detect BCL7B and key immune cell populations.

  • Focus particularly on B cells, dendritic cells, neutrophils, T cells, macrophages, and NK cells in BLCA, LGG, PRAD, and THCA samples, where significant correlations have been documented .

  • Quantify spatial relationships between BCL7B-expressing cells and infiltrating immune populations.

• Functional Mechanistic Studies:

  • Use BCL7B antibodies for immunoprecipitation followed by mass spectrometry to identify protein interaction partners in immune regulatory pathways.

  • Combine with proximity ligation assays to confirm specific in situ interactions with immune signaling components.

  • Design experiments targeting the cytokine-cytokine receptor interaction pathway and JAK/STAT signaling, which are correlated with BCL7B expression .

• Therapeutic Targeting Evaluation:

  • Utilize BCL7B antibodies to monitor expression changes following immune checkpoint blockade therapy.

  • Implement BCL7B immunostaining in pre- and post-treatment biopsies to assess potential as a response biomarker.

  • Consider developing therapeutic-grade antibodies if BCL7B proves to be a viable immunotherapy target.

By implementing this comprehensive approach, researchers can effectively utilize BCL7B antibodies to elucidate its specific contributions to immune evasion mechanisms and potential value as a therapeutic target or biomarker.