FAM187B Antibody

What is FAM187B and what are its known functions in human tissues?

FAM187B (Family with sequence similarity 187, member B) is a protein-coding gene also known as TMEM162 (Transmembrane protein 162) . The protein is identified by UniProt ID Q17R55 and Entrez Gene ID 148109 . While comprehensive functional studies on FAM187B are still emerging, it belongs to a family of proteins that are conserved among mammals. Current research indicates it is expressed in human tissues, though its specific cellular functions remain under investigation. Unlike the better-characterized FAM188B, which has been shown to enhance cell survival through interaction with USP7 and regulate p53 pathways , detailed pathway interactions for FAM187B are still being elucidated.

What types of FAM187B antibodies are available for research applications?

Several types of FAM187B antibodies are available for research purposes:

These antibodies are typically unconjugated and available in buffered aqueous glycerol solutions. Control fragments, such as the Human FAM187B (aa 219-306) recombinant protein, are also available for validation experiments .

What are the recommended applications for FAM187B antibodies?

FAM187B antibodies have been validated for several research applications:

• Immunohistochemistry (IHC): Used for detecting and localizing FAM187B in tissue sections with recommended dilutions ranging from 1:20 to 1:500 depending on the specific antibody .

• Immunofluorescence (IF): Useful for subcellular localization studies with recommended concentrations of 0.25-2 μg/mL for ICC-IF applications .

• ELISA: Validated for quantitative detection of FAM187B protein in various sample types .

These applications facilitate investigations into tissue expression patterns, protein localization, and quantitative analysis of FAM187B in experimental systems.

How should FAM187B antibodies be stored and handled for optimal performance?

For maximum stability and performance, FAM187B antibodies should be handled according to these guidelines:

• Storage temperature: Store at -20°C for long-term preservation .

• Buffer composition: Most commercial FAM187B antibodies are supplied in PBS with 0.02% sodium azide and 50% glycerol at pH 7.3 to maintain stability .

• Aliquoting: To prevent repeated freeze-thaw cycles, aliquot the antibody upon receipt if planning multiple uses over time.

• Shipping conditions: Typically shipped on wet ice; ensure proper storage immediately upon receipt .

• Stability: Most antibodies remain stable for at least one year when properly stored at the recommended temperature .

Always review the manufacturer's specific recommendations for each antibody product, as storage conditions may vary slightly between suppliers.

What validation methods should be used to confirm FAM187B antibody specificity?

Validating antibody specificity is crucial for generating reliable research data. For FAM187B antibodies, consider these validation approaches:

• Blocking experiments: Use recombinant FAM187B control fragments (such as the Human FAM187B aa 219-306 control fragment) at 100x molar excess relative to the antibody concentration. Pre-incubate the antibody-protein control fragment mixture for 30 minutes at room temperature before application to validate specificity .

• Orthogonal validation: Compare antibody-based detection with RNAseq or other non-antibody-based methods to confirm target expression patterns. Some commercial FAM187B antibodies have undergone orthogonal RNAseq validation .

• Negative controls: Include samples where the primary antibody is omitted to assess background signal and non-specific binding of the secondary antibody .

• Positive controls: Use tissues or cell lines with confirmed FAM187B expression, such as HeLa or MCF-7 cells, which have been used for validation of antibodies to related proteins .

• Western blot analysis: While not explicitly mentioned for all FAM187B antibodies in the search results, western blotting can verify antibody specificity by confirming the molecular weight of the detected protein.

How can researchers troubleshoot high background issues when using FAM187B antibodies in immunoassays?

High background signal is a common issue in antibody-based assays. To address this problem with FAM187B antibodies:

• Optimize blocking conditions: Increase blocking incubation time and consider using 5-10% normal serum from the same species as the detection antibody .

• Reduce primary antibody concentration: Dilute the antibody further to its optimal working concentration through titration experiments. For FAM187B antibodies, consider starting with the manufacturer's recommended dilutions (1:200-1:500 for IHC or 0.25-2 μg/mL for ICC-IF) and adjust as needed .

• Evaluate secondary antibody specificity: Run controls without primary antibody to check for non-specific secondary antibody binding. Ensure the secondary antibody is raised in a different species than your sample, and consider using pre-adsorbed secondary antibodies .

• Increase washing frequency and duration: Thoroughly wash between all steps to remove unbound antibodies that could contribute to background signal .

• For ELISA applications: If precipitates form in wells upon substrate addition, decrease substrate concentration. Read plates immediately after adding stop solution to avoid signal alteration .

• Substrate concentration: When using enzyme-conjugated systems, dilute the substrate and reduce incubation time if excessive signal is observed .

• Signal amplification adjustment: If using biotinylation or other signal amplification techniques, reduce the amplification level by decreasing the amount of biotin conjugated to the secondary antibody .

What control experiments are essential when using FAM187B antibodies?

To ensure reliable and interpretable results with FAM187B antibodies, implement these controls:

• Negative controls:

  • Omit primary antibody to assess secondary antibody non-specific binding

  • Use isotype controls (rabbit IgG at equivalent concentration) to evaluate non-specific binding

  • Include tissues or cells known not to express FAM187B

• Blocking controls:

  • Pre-incubate the antibody with FAM187B recombinant protein control fragment (aa 219-306) at 100x molar excess for 30 minutes at room temperature before application

• Positive controls:

  • Include tissues with confirmed FAM187B expression

  • Compare staining patterns with expression data from the Human Protein Atlas, which documents expression patterns across normal and cancer tissues

• Technical controls:

  • Include replicate samples to assess reproducibility

  • Evaluate multiple antibody dilutions to determine optimal signal-to-noise ratio

  • For quantitative applications, include standard curves using recombinant FAM187B protein

These controls help distinguish specific from non-specific signals and validate experimental findings.

How can researchers optimize immunohistochemistry protocols specifically for FAM187B detection?

Optimizing immunohistochemistry (IHC) protocols for FAM187B requires careful attention to several parameters:

• Tissue fixation and processing:

  • For formalin-fixed paraffin-embedded (FFPE) samples, optimize fixation time to preserve epitope accessibility

  • Consider epitope retrieval methods (heat-induced or enzymatic) to expose FAM187B epitopes that may be masked during fixation

• Antibody selection and dilution:

  • Begin with the manufacturer's recommended dilution range (1:200-1:500 for Sigma-Aldrich or 1:20-1:200 for SciCommHub products)

  • Perform a dilution series to determine optimal concentration for your specific tissue and fixation conditions

  • Consider polyclonal antibodies for increased sensitivity, particularly for proteins with low expression levels

• Blocking and incubation conditions:

  • Use 5-10% normal serum from the same species as the secondary antibody to minimize background

  • Optimize incubation time and temperature (typically overnight at 4°C or 1-2 hours at room temperature for primary antibody)

  • Consider using protein-free blocking buffers if high background persists

• Detection systems:

  • For low abundance targets, consider amplification systems such as tyramide signal amplification

  • Adjust substrate development time to optimize signal-to-noise ratio

  • For fluorescence detection, select fluorophores with spectral properties that minimize tissue autofluorescence interference

• Counterstaining and mounting:

  • Select counterstains that highlight cellular context without obscuring FAM187B staining

  • Use appropriate mounting media to preserve signal and reduce photobleaching for fluorescence applications

• Validation strategies:

  • Compare staining patterns with orthogonal methods such as in situ hybridization or RNAseq data

  • Document staining in multiple tissue types to establish specificity and expression patterns

What approaches can be used for multiplex detection of FAM187B alongside other proteins?

Investigating protein interactions and co-localization patterns requires multiplex detection approaches:

• Sequential immunostaining:

  • Perform successive staining rounds with complete stripping or inactivation of antibodies between rounds

  • Document signal after each round to distinguish overlapping signals

• Spectral unmixing techniques:

  • Use fluorophores with distinct spectral properties and apply spectral unmixing algorithms to separate overlapping emission spectra

  • Select secondary antibodies conjugated to fluorophores with minimal spectral overlap

• Primary antibody selection:

  • Choose primary antibodies raised in different host species to enable simultaneous detection with species-specific secondary antibodies

  • If using multiple rabbit antibodies (common for FAM187B), employ sequential staining with direct labeling of one antibody

• Proximity-based detection methods:

  • Utilize proximity ligation assays (PLA) to detect and visualize protein-protein interactions involving FAM187B

  • Consider FRET-based approaches to investigate molecular interactions below the diffraction limit

• Co-immunoprecipitation followed by immunoblotting:

  • Immunoprecipitate FAM187B and analyze co-precipitating proteins by mass spectrometry

  • This approach has been successful for related proteins like FAM188B, which was found to interact with p53 and USP7

• Antibody conjugation strategies:

  • Directly conjugate FAM187B antibodies to fluorophores, enzymes, or haptens to facilitate multiplexing

  • Consider zenon labeling or similar technologies for flexible antibody labeling

How can FAM187B antibodies be employed in advanced protein characterization studies?

Advanced characterization of FAM187B requires sophisticated methodological approaches:

• Protein interaction network analysis:

  • Perform immunoprecipitation followed by mass spectrometry to identify FAM187B-interacting proteins

  • Validate interactions using reciprocal co-immunoprecipitation with antibodies against putative binding partners

  • Consider using methodologies similar to those employed for FAM188B, which revealed interactions with important regulatory proteins such as p53 and USP7

• Post-translational modification (PTM) detection:

  • Use PTM-specific antibodies in combination with general FAM187B antibodies to investigate phosphorylation, ubiquitination, or other modifications

  • Employ phosphatase or deubiquitinase treatments prior to immunoblotting to confirm PTM specificity

• Subcellular localization studies:

  • Combine FAM187B immunofluorescence with markers for cellular compartments to precisely map subcellular distribution

  • Consider super-resolution microscopy techniques to resolve detailed localization patterns beyond the diffraction limit

• Functional antibody applications:

  • Explore techniques for antibody-mediated protein modulation similar to those described for selecting agonist antibodies through combinatorial antibody libraries

  • Consider intrabody approaches to manipulate FAM187B function in living cells

• Temporal dynamics investigation:

  • Use live-cell imaging with FAM187B antibody fragments to track protein dynamics

  • Apply FRAP (Fluorescence Recovery After Photobleaching) or related techniques to study FAM187B mobility and turnover

• Structural studies:

  • Employ conformation-specific antibodies to investigate structural states of FAM187B

  • Consider epitope mapping to identify antibody binding sites and structural domains

What strategies can be employed for developing novel FAM187B antibodies with enhanced properties?

Researchers seeking to develop improved FAM187B antibodies might consider these approaches:

• Epitope selection strategies:

  • Target unique, accessible regions of FAM187B with low homology to related proteins

  • Consider the immunogen sequence used in existing antibodies (e.g., DNFRLDEKTEFVWLDCPLGSMYRPVNWRANDTPLTWESQLSGQDFTTFLDPSTGGRQLQVFQPAVYKCFVQQELVAQFKPAASLETLE) as a starting point

  • Design peptides or recombinant fragments that specifically target functional domains

• Combinatorial library approaches:

  • Use phage display or similar technologies to select high-affinity antibodies from combinatorial libraries

  • Consider methodologies similar to those described for selecting agonist antibodies where lentiviral antibody libraries are used to infect eukaryotic cells with reporter systems

• Affinity maturation:

  • Perform directed evolution to enhance binding affinity of existing FAM187B antibodies

  • Utilize techniques such as error-prone PCR or CDR walking to generate variants with improved properties

• Validation enhancement:

  • Incorporate orthogonal validation methodologies similar to those used by established antibody providers (e.g., Prestige Antibodies)

  • Validate across multiple applications and sample types to ensure versatility

• Specialized antibody formats:

  • Develop recombinant antibody fragments (Fab, scFv) for applications requiring smaller molecular size

  • Create bispecific antibodies targeting FAM187B and interacting proteins for co-localization studies

• Enhanced specificity:

  • Perform negative selection against related family members to improve specificity

  • Consider cross-adsorption techniques to remove antibodies that recognize conserved epitopes

How can researchers investigate potential roles of FAM187B in disease processes?

Understanding FAM187B's potential role in disease requires strategic research approaches:

• Expression profiling:

  • Analyze FAM187B expression across normal and disease tissues using validated antibodies

  • Compare expression patterns with publicly available datasets and resources such as the Human Protein Atlas

• Genetic association studies:

  • Investigate potential links between FAM187B genetic variants and disease susceptibility

  • Correlate expression levels with genetic variations and disease phenotypes

• Functional genomics approaches:

  • Use CRISPR-Cas9 or RNAi technologies to modulate FAM187B expression and assess phenotypic consequences

  • Consider approaches similar to those used for FAM188B, where knockdown induced cell growth inhibition and increased apoptosis in cancer cell lines

• Protein interaction studies:

  • Identify FAM187B interaction partners through immunoprecipitation coupled with mass spectrometry

  • Investigate whether these interactions are altered in disease states

• Tissue and cellular phenotyping:

  • Examine cellular morphology, proliferation, apoptosis, and differentiation in response to FAM187B modulation

  • Correlate FAM187B expression with histopathological features in disease tissues

• Therapeutic targeting assessment:

  • Evaluate FAM187B as a potential therapeutic target if disease associations are established

  • Consider antibody-based therapeutic approaches if surface exposure is confirmed

What are the key considerations for integrating FAM187B antibody data with other -omics approaches?

Multi-omics integration provides comprehensive understanding of FAM187B biology:

• Correlative analysis frameworks:

  • Correlate protein expression data from FAM187B antibody studies with transcriptomic data

  • Integrate with genomic data to identify genetic variants that influence FAM187B expression or function

• Data normalization strategies:

  • Develop normalization approaches to compare antibody-based quantification with mass spectrometry-based proteomics

  • Consider batch effects and technical variations when integrating data from multiple platforms

• Pathway and network analysis:

  • Place FAM187B in the context of biological pathways using integrated protein-protein interaction data

  • Consider approaches similar to those used for FAM188B, which revealed connections to p53 pathway regulation

• Visualization tools:

  • Employ advanced visualization methods to represent multi-omics data integration

  • Use dimensionality reduction techniques to identify patterns across multiple data types

• Functional validation:

  • Design experiments to test hypotheses generated from integrated -omics analyses

  • Use antibody-based methods to validate predictions from computational analyses

• Public data repositories:

  • Contribute FAM187B antibody validation and expression data to public repositories

  • Leverage existing multi-omics datasets to generate hypotheses about FAM187B function