FAM24B Antibody

What is FAM24B protein and why is it studied?

FAM24B (Family with Sequence Similarity 24 Member B) is a human protein encoded by the FAM24B gene. It is also known by alternative names including AC073585.2, DKFZp667I0323, hypothetical protein LOC196792, and MGC45962 . While the specific function of FAM24B remains under investigation, researchers study this protein to understand its potential role in cellular processes and disease mechanisms. The protein contains specific amino acid sequences that are targeted by commercial antibodies, with a recombinant protein corresponding to the amino acid sequence TESCPALQCCEGYRMCASFDSLPPCCCDINEGL being used for antibody development .

What applications are FAM24B antibodies validated for?

Commercial FAM24B antibodies are primarily validated for immunological applications including:

• Immunohistochemistry (IHC) with recommended dilutions of 1:1000 - 1:2500

• Immunohistochemistry-Paraffin (IHC-P) with recommended dilutions of 1:1000 - 1:2500

• Enzyme-Linked Immunosorbent Assay (ELISA)

It's important to note that application-specific validation is critical, as antibody performance can vary significantly between different experimental contexts. Recent large-scale validation studies indicate that success in one application doesn't necessarily predict success in another, with immunofluorescence (IF) potentially being the best predictor of performance in Western blot (WB) and immunoprecipitation (IP) .

What types of FAM24B antibodies are commercially available?

Different types of FAM24B antibodies are available for research purposes:

When selecting a FAM24B antibody, consider that recent validation studies have demonstrated recombinant antibodies generally outperform both monoclonal and polyclonal antibodies across multiple applications, with success rates of 67% in Western blot, 54% in immunoprecipitation, and 48% in immunofluorescence compared to lower rates for monoclonal and polyclonal alternatives .

How should I validate a newly purchased FAM24B antibody?

Robust validation of FAM24B antibodies is essential before conducting experiments. A comprehensive validation approach includes:

• Knockout (KO) validation: The gold standard approach involves testing the antibody on samples where FAM24B has been genetically knocked out. This method provides definitive evidence of specificity by comparing signal between wild-type and KO samples .

• Specificity testing: Verify that the antibody detects only FAM24B by examining if the signal disappears in samples lacking the target protein. Large-scale validation studies have employed protein arrays containing the target protein plus hundreds of non-specific proteins to verify specificity .

• Application-specific validation: Test the antibody in your specific application of interest. Success in one application doesn't guarantee success in another. Validation studies have shown that only about 27% of polyclonal, 41% of monoclonal, and 67% of recombinant antibodies successfully detect their targets in Western blot .

• Positive controls: Use recombinant FAM24B protein as a positive control. Commercial recombinant FAM24B proteins covering amino acids 22-94 with His-tags are available for this purpose .

What are the recommended experimental conditions for FAM24B antibody applications?

When designing experiments with FAM24B antibodies, consider these application-specific recommendations:

For Immunohistochemistry (IHC and IHC-P):

• Dilution range: 1:1000 - 1:2500

• Storage conditions: Store at 4°C short term; aliquot and store at -20°C for long term

• Avoid freeze-thaw cycles to maintain antibody integrity

• Buffer system: PBS (pH 7.2) with 40% Glycerol and 0.02% Sodium Azide

Always optimize these conditions for your specific experimental system, as factors like tissue fixation methods, antigen retrieval techniques, and detection systems can significantly impact antibody performance.

What controls should I implement when working with FAM24B antibodies?

Implementing proper controls is crucial for meaningful interpretation of FAM24B antibody results:

• Negative controls:

  • Isotype control: Use matched isotype IgG from the same species

  • Secondary antibody only: Omit primary antibody to detect non-specific binding

  • FAM24B knockout or knockdown samples: Ideal for confirming specificity

• Positive controls:

  • Tissues or cells known to express FAM24B

  • Recombinant FAM24B protein

  • Overexpression systems: Cells transfected to overexpress FAM24B

• Technical replicates: Include multiple technical replicates to assess reproducibility of findings

Validation studies have demonstrated that proper controls can identify significant issues with antibody specificity, with more than 50% of tested commercial antibodies failing in one or more applications .

How do different types of FAM24B antibodies compare in performance?

When selecting a FAM24B antibody, understanding performance differences between antibody types is critical:

Recent large-scale validation studies clearly demonstrate that recombinant antibodies significantly outperform both monoclonal and polyclonal antibodies across multiple applications . If available, recombinant FAM24B antibodies would likely provide the most reliable results for research applications.

What methodological approaches can improve FAM24B antibody performance?

Optimizing experimental protocols can significantly enhance FAM24B antibody performance:

• Epitope retrieval optimization: For IHC-P applications, systematically test different antigen retrieval methods (heat-induced vs. enzymatic, different pH buffers) to maximize epitope accessibility.

• Signal amplification strategies: Consider tyramide signal amplification or polymeric detection systems when working with low-abundance targets.

• Blocking optimization: Test different blocking agents (BSA, normal serum, commercial blockers) to minimize background signal while preserving specific binding.

• Computer-aided antibody design: Recent advances in computational antibody design combine physics-based and AI-driven methods to improve antibody performance characteristics while maintaining target binding . These approaches have demonstrated success in:

  • Traversing sequence landscapes to identify diverse binders

  • Rescuing binding from escape mutations

  • Improving developability characteristics while preserving binding properties

• Validation sequence: Consider that success in immunofluorescence has been found to be the best predictor of performance in Western blot and immunoprecipitation, suggesting IF could be used as an initial screening method before proceeding to other applications .

What factors influence FAM24B antibody specificity and sensitivity?

Multiple factors affect the specificity and sensitivity of FAM24B antibodies:

• Epitope accessibility: The conformation of FAM24B in your experimental system affects antibody binding. The FAM24B antibody was developed against a specific amino acid sequence (TESCPALQCCEGYRMCASFDSLPPCCCDINEGL), and accessibility of this sequence varies across applications .

• Post-translational modifications: These can mask epitopes or create new ones, affecting antibody recognition.

• Protein interactions: FAM24B interactions with other proteins may sterically hinder antibody binding.

• Sample preparation: Denaturation, fixation, and embedding methods significantly impact epitope presentation.

• Antibody format: The format (whole IgG, Fab, scFv) affects tissue penetration and background binding.

Large-scale validation studies have revealed that even widely used commercial antibodies may perform poorly when rigorously tested, with hundreds of underperforming antibodies being used in numerous published studies . This highlights the critical importance of thorough validation before experimental use.

How do I address non-specific binding with FAM24B antibodies?

Non-specific binding is a common challenge when working with FAM24B antibodies. Implement these strategies to minimize background:

• Optimize blocking conditions: Test different blocking agents (BSA, normal serum from the same species as the secondary antibody, commercial blockers) and concentrations.

• Titrate antibody concentration: Perform a dilution series to identify the optimal antibody concentration that maximizes specific signal while minimizing background.

• Increase washing stringency: Add detergents (Tween-20, Triton X-100) to wash buffers and increase wash duration/frequency.

• Pre-absorb antibody: Incubate with irrelevant tissue lysate to remove antibodies that bind non-specifically.

• Validate antibody specificity: Confirm specificity using knockout or knockdown controls. Studies have shown that many commercial antibodies exhibit non-specific binding that is only detectable when properly controlled experiments are performed .

What should I do if my FAM24B antibody shows inconsistent results between experiments?

Inconsistent results with FAM24B antibodies may stem from several sources:

• Antibody degradation: Store according to manufacturer recommendations (4°C short-term; -20°C long-term aliquots) . Avoid freeze-thaw cycles.

• Batch variation: Particularly relevant for polyclonal antibodies. Consider switching to recombinant antibodies, which demonstrate higher consistency between batches .

• Protocol standardization: Document and standardize every aspect of your protocol:

  • Sample collection and processing

  • Buffer compositions

  • Incubation times and temperatures

  • Detection methods

• Sample heterogeneity: Inconsistent expression levels of FAM24B across samples can produce variable results.

• Experimental validation: Implement the validation approaches described in recent literature, including side-by-side comparisons using knockout controls, which have proven effective in identifying reliable antibodies .

How can I determine if published research using FAM24B antibodies is reliable?

When evaluating published research utilizing FAM24B antibodies, consider these assessment criteria:

• Validation methods: Look for proper validation using knockout/knockdown controls, recombinant expression systems, or orthogonal detection methods.

• Antibody identification: Check if complete antibody information is provided (manufacturer, catalog number, lot number, RRID).

• Control implementation: Assess the quality and appropriateness of experimental controls.

• Methodology details: Evaluate if sufficient experimental details are provided to reproduce the results.

• Multiple antibody verification: Studies using multiple antibodies targeting different FAM24B epitopes provide stronger evidence.

Recent validation initiatives have revealed that many antibodies used in published literature perform poorly when rigorously tested. One study found that hundreds of underperforming antibodies identified in systematic validation had been used in numerous published articles, raising concerns about result reliability . This underscores the importance of critical evaluation when consulting published FAM24B antibody research.

What emerging technologies might improve FAM24B antibody development and validation?

Several innovative approaches are advancing antibody research that may benefit FAM24B studies:

• AI-driven antibody design: Computational pipelines incorporating physics-based and machine learning methods can generate candidate antibodies with improved binding and developability characteristics . These approaches have demonstrated success in:

  • Identifying sequence-diverse antibodies that retain binding

  • Rescuing binding from escape mutations

  • Improving developability while maintaining binding properties

• High-throughput validation platforms: Scalable antibody validation procedures using genetic knockout cell lines provide definitive specificity assessment . These systems can:

  • Test hundreds of antibodies simultaneously

  • Validate across multiple applications (WB, IP, IF)

  • Generate public data repositories to guide antibody selection

• Antibody engineering: Advanced protein engineering techniques can enhance FAM24B antibody performance by improving:

  • Specificity through directed evolution

  • Stability through framework optimization

  • Affinity through targeted mutagenesis

• Renewable antibody resources: Development of recombinant antibodies provides consistent, renewable reagents that outperform traditional antibody formats .

How can computational approaches enhance FAM24B antibody research?

Computational methods are increasingly valuable for antibody research:

• Structure-based antibody design: Using computational modeling to predict antibody-antigen interactions and design improved binding interfaces .

• Machine learning for property prediction: AI systems can predict antibody developability characteristics and binding properties from sequence data .

• Bayesian optimization: This approach can guide experimental testing through multiple rounds of design and validation to efficiently optimize antibody properties .

• Inverse folding models: These computational approaches can help restore binding activity after antigen mutations, potentially important for adapting FAM24B antibodies to different experimental contexts .

Recent studies have demonstrated that combining computational prediction with focused experimental validation can dramatically reduce the resources required for developing high-quality antibodies while improving success rates .

What standardized validation criteria should be applied to FAM24B antibodies?

To advance FAM24B antibody research quality, these standardized validation criteria should be widely adopted:

• Application-specific validation: Antibodies should be validated specifically for each application (WB, IP, IF, IHC) rather than extrapolating performance between methods .

• Genetic knockout controls: Testing against samples lacking FAM24B expression provides definitive specificity assessment .

• Standardized reporting: Complete documentation of antibody characteristics, validation methods, and experimental conditions enables reproducibility.

• Independent validation: Third-party assessment rather than relying solely on manufacturer claims .

• Data sharing: Contributing validation results to public repositories advances community knowledge.

Studies suggest that implementing these standardized validation practices could save a significant portion of the estimated $1 billion wasted annually on research involving ineffective antibodies, while dramatically improving research reliability .