FAM153C Antibody

What is FAM153C and what cellular processes is it involved in?

FAM153C (Family with Sequence Similarity 153, Member C) is a protein coding gene with implications in cell growth, differentiation, and potentially tumorigenesis. While not extensively characterized, its role in these fundamental cellular processes makes it a promising target for studies in cancer research and developmental biology . The protein is identified by UniProt Code Q494X1 and is also known as NY-REN-7-like according to some databases .

Current research suggests FAM153C may be involved in regulatory pathways that influence cell proliferation, though the specific mechanisms remain under investigation. Understanding FAM153C function and regulation could provide valuable insights into disease mechanisms and potential therapeutic interventions .

What expression patterns of FAM153C have been observed in human tissues?

Based on immunohistochemistry data from available antibodies, FAM153C expression has been detected in several human tissues. Notable expression has been observed in adrenal gland tissue, as demonstrated by immunohistochemistry of paraffin-embedded samples using antibodies at dilutions of 1:100 . Additionally, placental tissue has been used as a sample source for antibody validation, suggesting expression in this tissue type as well .

The protein appears to have varying expression levels across different tissue types, though comprehensive tissue expression profiles are still being developed as more research groups incorporate FAM153C analysis into their studies.

What criteria should be used when selecting a FAM153C antibody for research applications?

When selecting a FAM153C antibody for research, consider the following critical parameters:

For Western blot applications, antibodies that recognize the human FAM153C protein with high specificity are recommended. Some antibodies have been validated with specific dilution ranges (e.g., 1:1000-2000 for Western blot applications) .

How should FAM153C antibodies be validated before experimental use?

A comprehensive validation protocol for FAM153C antibodies should include:

• Positive Control Selection: Use tissues/cell lines known to express FAM153C (e.g., placenta lysates, adrenal gland tissue)

• Western Blot Validation:

  • Run both positive control and experimental samples

  • Check for bands at the expected molecular weight (~12.7 kDa for full-length protein)

  • Include negative controls (tissues/cells with no or low FAM153C expression)

  • Perform peptide competition assays to confirm specificity

• IHC Validation:

  • Test on known positive tissues (e.g., adrenal gland)

  • Optimize antibody dilution (starting with manufacturer recommendations, such as 1:20-1:200)

  • Include appropriate negative controls (omitting primary antibody)

  • Consider antigen retrieval methods if needed

• Specificity Controls:

  • siRNA knockdown of FAM153C to confirm signal reduction

  • Comparison with alternative antibodies targeting different epitopes

  • Peptide blocking experiments

Each new lot of antibody should undergo basic validation to ensure consistent performance in your experimental system.

What are the optimal conditions for using FAM153C antibodies in Western blot applications?

For optimal Western blot results with FAM153C antibodies:

Sample Preparation and Protocol:

• Extract total protein using standard lysis buffers (e.g., RIPA buffer with protease inhibitors)

• Quantify protein and load 20-40 μg per lane

• Separate proteins using SDS-PAGE (10-15% gel recommended for this lower molecular weight protein)

• Transfer to PVDF or nitrocellulose membrane (PVDF may provide better results for lower abundance proteins)

• Block with 5% non-fat milk or BSA in TBST (1 hour at room temperature)

• Incubate with primary FAM153C antibody at optimized dilution (typically 1:1000-2000) overnight at 4°C

• Wash 3× with TBST

• Incubate with appropriate HRP-conjugated secondary antibody (anti-rabbit for most FAM153C antibodies)

• Develop using chemiluminescence detection

Critical Considerations:

• Expected molecular weight: ~12.7 kDa for the native protein

• Some FAM153C antibodies may require BSA rather than milk for blocking

• Longer exposure times may be needed for detection if expression is low

• Consider including positive control samples (e.g., placenta lysate)

What immunohistochemistry protocols are recommended for FAM153C detection in tissue samples?

For optimal IHC detection of FAM153C in tissue samples:

IHC Protocol for FAM153C:

• Prepare paraffin-embedded tissue sections (4-6 μm thickness)

• Deparaffinize and rehydrate sections using standard protocols

• Perform heat-induced epitope retrieval (HIER) using citrate buffer (pH 6.0) or EDTA buffer (pH 9.0)

• Block endogenous peroxidase activity with 3% H₂O₂

• Block non-specific binding with 5-10% normal serum

• Incubate with primary FAM153C antibody at optimized dilution (start with 1:100 as demonstrated in adrenal gland tissue)

• Incubate overnight at 4°C or 1-2 hours at room temperature

• Wash with PBS or TBS

• Apply appropriate secondary antibody and detection system

• Counterstain, dehydrate, and mount

Important Considerations:

• Pilot studies with different dilutions (1:20-1:200) are recommended to optimize signal-to-noise ratio

• Adrenal gland tissue serves as a good positive control for FAM153C detection

• Include appropriate negative controls (omission of primary antibody, isotype control)

• Document all staining patterns observed, as the exact subcellular localization of FAM153C may vary between tissue types

How can FAM153C antibodies be utilized in cancer research studies?

FAM153C has potential implications in tumorigenesis, making antibodies against this protein valuable tools in cancer research:

Research Applications in Oncology:

• Expression Profiling: Quantify FAM153C expression across cancer types and correlate with clinical outcomes

  • IHC on tissue microarrays (TMAs) using optimized dilutions (1:20-1:200)

  • Western blot analysis of cancer cell lines and tumor samples

• Functional Studies:

  • Combine with knockdown/overexpression approaches to assess functional consequences

  • Correlate expression with cell proliferation, migration, and invasion assays

• Mechanistic Investigations:

  • Co-immunoprecipitation to identify protein interaction partners

  • Chromatin immunoprecipitation if nuclear localization is confirmed

  • Immunofluorescence to determine subcellular localization in cancer versus normal cells

• Prognostic Biomarker Development:

  • Evaluate FAM153C as a potential biomarker using validated antibodies on patient cohorts

  • Correlate expression patterns with clinical data similar to approaches used for other cancer-associated genes

Emerging research suggests potential roles of FAM153C in cellular pathways relevant to cancer biology, warranting further investigation using well-validated antibodies .

What approaches can be used to investigate potential binding partners or complexes involving FAM153C?

To identify and characterize FAM153C protein interactions:

Methodological Approaches:

• Co-Immunoprecipitation (Co-IP):

  • Use FAM153C antibodies for immunoprecipitation from cell lysates

  • Identify binding partners through mass spectrometry

  • Validate interactions with reverse Co-IP using antibodies against identified partners

  • Recommended antibody dilution: 1:50-1:200 for immunoprecipitation

• Proximity Ligation Assay (PLA):

  • Combine FAM153C antibody with antibodies against suspected interaction partners

  • Visualize protein-protein interactions in situ with subcellular resolution

  • Requires high antibody specificity and appropriate controls

• Pull-down Assays with Recombinant Protein:

  • Use recombinant FAM153C protein (like the Myc-DYKDDDDK tagged version)

  • Pull down potential interacting partners from cell lysates

  • Identify through Western blot or mass spectrometry

• Yeast Two-Hybrid Screening:

  • Use as a complementary approach to antibody-based methods

  • Validate hits using antibody-based approaches

When investigating potential roles in DNA repair pathways or complexes similar to those studied in homologous recombination screens , additional controls and specialized approaches may be needed to confirm functional interactions.

What are common challenges when working with FAM153C antibodies and how can they be addressed?

Storage and Handling Recommendations:

• Store antibody at -20°C for long-term storage or according to manufacturer specifications

• Avoid repeated freeze-thaw cycles (limit to 2-3 cycles)

• Aliquot antibodies upon receipt to minimize freeze-thaw damage

• Use proper concentrations of preservatives if diluting for storage

How can antibody-based detection of FAM153C be integrated with other molecular approaches for comprehensive analysis?

For multidimensional characterization of FAM153C:

• Integrated Genomic and Proteomic Analysis:

  • Combine antibody-based protein detection with RNA expression analysis

  • Correlate protein levels (by Western blot/IHC) with mRNA expression (by qRT-PCR)

  • Investigate potential discrepancies between transcriptional and post-transcriptional regulation

• Functional Genomics Integration:

  • Use FAM153C antibodies to validate siRNA/CRISPR knockdown efficiency

  • Combine with phenotypic assays to correlate protein levels with functional outcomes

  • Potential application in homologous recombination screens similar to those described for other genes

• Single-Cell Analysis:

  • Use FAM153C antibodies for flow cytometry or mass cytometry (CyTOF)

  • Combine with other markers to characterize heterogeneity within cell populations

  • Optimize antibody conditions specifically for flow applications

• Multi-omics Data Integration:

  • Correlate FAM153C protein levels with genomic alterations (copy number, mutations)

  • Apply computational methods similar to those used in transcriptomic studies

  • Consider topologically associated domain analysis when studying regulatory relationships

• Translational Research Applications:

  • Validate antibody performance in clinically relevant sample types

  • Develop standardized protocols applicable across research sites

  • Consider automated image analysis for quantitative IHC assessment

This integrated approach provides a comprehensive view of FAM153C biology beyond what can be achieved with antibody-based detection alone.

What is the potential significance of FAM153C in disease mechanisms based on current research?

While research on FAM153C is still emerging, several lines of evidence suggest potential significance in disease mechanisms:

• Cancer Biology: Preliminary data suggests FAM153C may have implications in tumorigenesis through its role in cell growth and differentiation processes . Similar to other protein family members studied in cancer contexts, dysregulation of FAM153C may contribute to oncogenic pathways.

• Cell Cycle Regulation: Based on patterns observed with related proteins studied in cancer research, FAM153C may participate in pathways involving cell cycle control, which has significant implications for both developmental processes and disease states .

• Cellular Stress Response: Some research suggests potential involvement in cellular response pathways, though specific mechanisms require further investigation with validated antibodies and functional studies.

• Potential Therapeutic Target: If functional studies confirm roles in disease-relevant pathways, FAM153C could represent a novel therapeutic target, particularly in cancers where expression is dysregulated.

Future research using well-validated antibodies will be essential to elucidate the precise functional roles of FAM153C in normal physiology and disease states.

What emerging technologies might enhance FAM153C research beyond traditional antibody applications?

Emerging technologies that could advance FAM153C research include:

• CRISPR-Based Tagging:

  • Endogenous tagging of FAM153C to avoid antibody specificity issues

  • Live-cell imaging of tagged protein for dynamic localization studies

  • Reduces reliance on antibody validation for certain applications

• Proximity-Based Labeling Techniques:

  • BioID or APEX2 fusion proteins to identify proximal proteins

  • Maps the protein interaction neighborhood without relying on stable interactions

  • Complements traditional antibody-based co-IP approaches

• Super-Resolution Microscopy:

  • Combining highly specific FAM153C antibodies with super-resolution techniques

  • Reveals detailed subcellular localization beyond conventional microscopy limits

  • Potential to identify previously unrecognized functional compartmentalization

• Single-Cell Proteomics:

  • Analysis of FAM153C expression at single-cell resolution

  • Reveals heterogeneity not captured by bulk analysis techniques

  • May identify rare cell populations with distinctive FAM153C expression patterns

• Spatial Transcriptomics Integration:

  • Combining antibody-based protein detection with spatial transcriptomics

  • Maps both protein expression and associated transcriptional programs

  • Particularly valuable for tissue-based studies and understanding cellular microenvironments

These approaches can complement traditional antibody applications to provide more comprehensive insights into FAM153C biology and disease relevance.