ITGB1BP2 Antibody

What is ITGB1BP2 and why is it significant in research?

ITGB1BP2, also known as Melusin 2, is an integrin beta 1 binding protein with a calculated molecular weight of approximately 38 kDa, though it is typically observed at 40-45 kDa in experimental conditions . This protein plays significant roles in cardiac and muscle tissue biology, making it an important target for cardiovascular and musculoskeletal research. The antibodies targeting this protein are essential tools for investigating its expression, localization, and functional relationships in various experimental models and human tissues.

What types of ITGB1BP2 antibodies are available for research applications?

Based on the search results, researchers have access to several types of ITGB1BP2 antibodies with different characteristics:

• Polyclonal antibodies: Such as ABIN204995 (rabbit-derived, targeting N-term) and 21359-1-AP (rabbit-derived)

• Monoclonal antibodies: Including those targeting specific amino acid regions like AA 246-319

• Conjugated and non-conjugated formats: Most common are unconjugated, but HRP-conjugated options exist for certain applications

These antibodies vary in their targeting epitopes (N-terminal, C-terminal, or specific amino acid sequences), making them suitable for different experimental needs and protein conformations .

How should I select the appropriate ITGB1BP2 antibody for my specific research model?

When selecting an ITGB1BP2 antibody, consider these critical factors:

• Species reactivity: Different antibodies show specific reactivity patterns. For example, ABIN204995 reacts with human, mouse, rat, rabbit, bat, and monkey samples , while 21359-1-AP demonstrates reactivity with human, mouse, and rat samples .

• Application compatibility: Determine whether your intended application (WB, IHC, ELISA, IF/ICC) is validated for the antibody. For instance, 21359-1-AP is validated for WB (1:500-1:1000 dilution) and IF/ICC (1:10-1:100 dilution) .

• Epitope targeting: Consider whether the antibody's epitope (N-terminal vs. C-terminal) is accessible in your experimental conditions, especially if you're working with protein fragments or in conditions that might affect protein conformation .

• Validation data: Review existing validation data for your tissue or cell type of interest before making a selection .

What are the optimal sample preparation techniques for different ITGB1BP2 antibody applications?

Sample preparation varies by application:

For Western Blotting:

• Use fresh tissue samples (heart tissue shows good ITGB1BP2 expression)

• Follow standard protein extraction protocols with protease inhibitors

• Protein denaturation conditions should be optimized as ITGB1BP2's observed molecular weight (40-45 kDa) differs slightly from its calculated weight (38 kDa)

For Immunofluorescence/ICC:

• HepG2 cells have shown positive results with 21359-1-AP

• Standard fixation protocols (4% paraformaldehyde) are suitable

• Permeabilization and blocking steps are critical for reducing background

For Immunohistochemistry:

• Both fresh-frozen and paraffin-embedded sections can be used

• Antigen retrieval may be necessary for optimal epitope accessibility, especially for paraffin sections

How can I optimize ITGB1BP2 antibody performance for challenging tissues or cell types?

For challenging samples, consider these optimization strategies:

• Dilution optimization: Though recommended dilutions exist (e.g., 1:500-1:1000 for WB with 21359-1-AP ), systematic titration experiments are necessary for each specific experimental system.

• Signal enhancement techniques:

  • For WB: Extended exposure times, more sensitive detection systems

  • For IHC/IF: Amplification systems (tyramide, polymer-based)

  • Increased antibody incubation times at lower temperatures (e.g., overnight at 4°C)

• Background reduction strategies:

  • More stringent washing procedures

  • Optimized blocking buffers (testing BSA vs. serum-based blockers)

  • Pre-adsorption with relevant tissues if cross-reactivity is suspected

• Sample-specific protocols: As noted in the antibody documentation, "Sample-dependent, Check data in validation data gallery" , indicating the need for optimization based on specific sample types.

What are the most common technical challenges when working with ITGB1BP2 antibodies and how can they be resolved?

Common challenges include:

• Multiple banding patterns in Western blots:

  • Possible post-translational modifications or isoforms

  • Solution: Use positive control tissues (human heart) with validated antibody dilutions

  • Verify with alternative antibodies targeting different epitopes

• Low signal intensity:

  • Increase protein loading (for WB) or antibody concentration

  • Optimize antigen retrieval methods for IHC/IF

  • Consider enhanced detection systems

• Non-specific binding:

  • Perform careful blocking optimization

  • Include appropriate negative controls (isotype controls, secondary-only controls)

  • Consider using tissue from knockout models if available

• Cross-reactivity with unexpected species:

  • Review BLAST analysis data provided with antibodies. For example, ABIN204995 shows 100% identity with mouse, rat, rabbit and 92% with dog, bovine, horse

How should I validate ITGB1BP2 antibody specificity for my experiments?

Rigorous validation should include:

• Multiple detection methods:

  • Compare results across different applications (WB, IHC, IF) when possible

  • Use antibodies targeting different epitopes of ITGB1BP2 (N-terminal vs. C-terminal)

• Appropriate controls:

  • Positive controls: Human heart tissue has demonstrated reliable detection

  • Negative controls: Tissues or cells known not to express ITGB1BP2

  • Peptide competition assays with the immunizing peptide

  • siRNA knockdown or CRISPR knockout validation when feasible

• Expected molecular weight verification:

  • ITGB1BP2 should appear at 40-45 kDa in Western blots, slightly higher than its calculated 38 kDa weight

  • Check for appropriate subcellular localization patterns in IF/IHC

• Cross-reference with published literature and antibody validation repositories

What approach should I take when ITGB1BP2 antibody results conflict with other data or published findings?

When facing contradictory results:

• Technical verification:

  • Repeat experiments with freshly prepared reagents

  • Test multiple antibody lots or sources

  • Verify experimental conditions match those in referenced studies

• Biological explanations:

  • Consider cell/tissue-specific expression patterns

  • Evaluate potential post-translational modifications affecting epitope accessibility

  • Examine potential splice variants or isoforms

• Methodological differences:

  • Analyze differences in sample preparation, fixation, or extraction methods

  • Compare antibody clones, epitopes, and host species used in conflicting studies

  • Evaluate differences in detection methods or sensitivity

• Confirming results with orthogonal techniques:

  • mRNA expression analysis (qPCR, RNA-seq)

  • Mass spectrometry-based proteomics

  • Functional assays to correlate with antibody-based detection

How can ITGB1BP2 antibodies be incorporated into multiplexed detection systems?

Strategies for multiplexed analysis include:

• Multi-color immunofluorescence:

  • Combining ITGB1BP2 detection with other markers of interest

  • Requires careful selection of secondary antibodies with minimal spectral overlap

  • Consider sequential detection protocols if antibody host species are limiting

• Co-immunoprecipitation studies:

  • ITGB1BP2 antibodies can be used to pull down protein complexes

  • Can reveal interaction partners and complex formation

  • Requires validation of antibody efficacy in non-denaturing conditions

• Tissue microarray analysis:

  • High-throughput screening of ITGB1BP2 expression across multiple samples

  • Allows for standardized comparison across tissue types or disease states

  • Requires optimization of IHC conditions for consistent results

• Flow cytometry applications:

  • Though not specifically mentioned in the search results, protocols could be optimized for flow cytometric detection in suitable cell types

What are the considerations for quantitative analysis of ITGB1BP2 using antibody-based detection methods?

For quantitative applications:

• Standardization approaches:

  • Use calibrated protein standards for Western blot quantification

  • Include internal loading controls (housekeeping proteins) for normalization

  • Prepare standard curves with recombinant ITGB1BP2 protein for ELISA applications

• Image analysis for IHC/IF:

  • Standardized acquisition parameters (exposure time, gain settings)

  • Appropriate background subtraction methods

  • Computer-assisted quantification of staining intensity and localization

• Quantitative ELISA techniques:

  • Sandwich ELISA methods are available for mouse samples (e.g., ABIN6228935)

  • Ensure proper sample dilution to stay within the linear range of detection

  • Include technical replicates and inter-assay controls

• Statistical considerations:

  • Power analysis to determine appropriate sample sizes

  • Appropriate statistical tests for experimental design

  • Methods for handling outliers and non-normal distributions