SCGB1A1 Recombinant Monoclonal Antibody

What is SCGB1A1 and what are its primary biological functions?

SCGB1A1, also known as Clara cell secretory protein (CCSP), is a protein primarily produced by Clara cells in the respiratory tract. Its main biological functions include protecting the lungs from oxidative stress, inflammation, and pathogens, while also contributing to lung homeostasis and repair processes . The protein is relatively small, with a reported molecular mass of approximately 10 kilodaltons . SCGB1A1 is also known by several other names in the scientific literature, including CC10, CC16, uteroglobin, UP1, and CCPBP, which may reflect its various biological roles in different tissues and physiological contexts .

Beyond its pulmonary functions, SCGB1A1 has been implicated in immunomodulatory processes and may play roles in conditions ranging from asthma to lung cancer. Its expression patterns change in response to various pathological conditions, making it an important biomarker in respiratory research and potentially in clinical applications.

How are SCGB1A1 recombinant monoclonal antibodies generated?

SCGB1A1 recombinant monoclonal antibodies are generated through sophisticated in vitro expression systems. The process begins with the immunization of rabbits using synthesized peptides derived from human SCGB1A1 protein. Subsequently, SCGB1A1 antibody DNA sequences are cloned from these immunoreactive rabbits .

The production process continues with the insertion of genes encoding the SCGB1A1 antibodies into plasmid vectors. These recombinant plasmid vectors are then transfected into host cells to facilitate antibody expression . Following expression, the antibodies undergo affinity-chromatography purification to ensure high purity and specificity. Before commercial release, these antibodies are thoroughly tested for functionality in applications such as ELISA and flow cytometry, specifically evaluating their reactivity with human SCGB1A1 protein .

Some SCGB1A1 antibodies are produced using purified recombinant proteins as immunogens. For example, some manufacturers use recombinant Mouse Uteroglobin/SCGB1A1 (specifically from UniProt Q06318; Met1-Phe96) to develop rabbit monoclonal antibodies with precise specificity .

What are the common applications for SCGB1A1 recombinant monoclonal antibodies?

SCGB1A1 recombinant monoclonal antibodies support a wide range of experimental techniques in respiratory biology, immunology, and molecular research. Based on manufacturer specifications, these antibodies demonstrate utility across multiple applications:

The versatility of these antibodies allows researchers to perform comprehensive analyses of SCGB1A1 expression, localization, and function across different experimental systems . This multi-application capability is particularly valuable for validating findings across different methodological platforms.

What species reactivity can be expected with SCGB1A1 antibodies?

Species reactivity is a critical consideration when selecting SCGB1A1 antibodies for research. Based on the available information, most SCGB1A1 recombinant monoclonal antibodies demonstrate reactivity with human samples, while some exhibit cross-reactivity with mouse and rat orthologs .

The search results indicate that commercially available SCGB1A1 antibodies vary in their species reactivity profiles:

When selecting an antibody for multi-species studies, researchers should carefully verify the cross-reactivity profile through manufacturer validation data or preliminary testing. Some antibodies specifically show no cross-reactivity with human SCGB1A1 despite strong reactivity with mouse samples, as noted in the Bio-Techne product specifications .

What is the difference between polyclonal and monoclonal SCGB1A1 antibodies?

The fundamental difference between polyclonal and monoclonal SCGB1A1 antibodies lies in their epitope recognition and production methodology. Monoclonal antibodies recognize a single epitope on the SCGB1A1 protein and are produced from a single B cell clone, ensuring consistent lot-to-lot reproducibility. In contrast, polyclonal antibodies recognize multiple epitopes and are derived from different B cell populations.

For SCGB1A1 research, this distinction has several important implications:

Recombinant monoclonal antibodies represent a further advancement, combining the specificity of monoclonals with recombinant production technology that enhances reproducibility and potentially reduces animal use in antibody production .

How does antibody selection affect SCGB1A1 detection sensitivity in different tissue types?

The selection of appropriate SCGB1A1 antibodies significantly influences detection sensitivity across different tissue types due to several tissue-specific factors. SCGB1A1 expression levels vary substantially between tissues, with highest expression in lung Clara cells but also presence in other epithelial tissues. This variable expression necessitates careful antibody selection.

Tissue-specific factors affecting antibody performance include:

• Tissue fixation effects on epitope accessibility - formalin fixation can mask SCGB1A1 epitopes differently in various tissues

• Endogenous peroxidase activity levels - particularly relevant in lung tissues where SCGB1A1 is primarily expressed

• Lipid content variations - affecting penetration and nonspecific binding

• Background autofluorescence - especially in lung tissue with elastin content

Researchers should consider conducting preliminary titration experiments with different antibody concentrations across tissue types to determine optimal sensitivity thresholds. For lung tissue, where SCGB1A1 is abundantly expressed, lower antibody concentrations may provide sufficient sensitivity, while other tissues might require higher concentrations or signal amplification methods .

Validation with multiple detection methods is strongly recommended when studying SCGB1A1 across different tissues to confirm specificity and rule out potential false positives due to cross-reactivity with related secretoglobin family members.

What factors should be considered when optimizing SCGB1A1 antibody dilution for immunohistochemistry?

Optimizing SCGB1A1 antibody dilution for immunohistochemistry requires systematic evaluation of multiple parameters to achieve the ideal balance between specific signal and background. Based on manufacturer recommendations, typical dilution ranges for SCGB1A1 antibodies in IHC applications are between 1:100-1:500 .

Key factors to consider during optimization include:

A systematic approach to optimization involves testing a dilution series (e.g., 1:100, 1:200, 1:400) while keeping other variables constant. Visual assessment of signal-to-noise ratio across multiple samples and tissue types will help determine the optimal working dilution . The specificity of staining should be confirmed using appropriate positive controls (lung tissue) and negative controls (tissues known not to express SCGB1A1).

How can researchers validate the specificity of SCGB1A1 antibodies in their experimental system?

Validation of SCGB1A1 antibody specificity is crucial for ensuring reliable research outcomes. A comprehensive validation strategy should employ multiple complementary approaches:

• Positive and Negative Control Tissues:

  • Positive controls: Normal lung tissue (Clara cells should show strong positivity)

  • Negative controls: Tissues without SCGB1A1 expression or SCGB1A1 knockout models

• Peptide Competition Assay:

  • Pre-incubate the antibody with excess SCGB1A1 recombinant protein or immunizing peptide

  • Specific staining should be abolished or significantly reduced

• Orthogonal Method Validation:

  • Confirm protein expression using alternative techniques (qPCR, mass spectrometry)

  • Compare results across different antibody clones targeting different SCGB1A1 epitopes

• Western Blot Analysis:

  • Verify correct molecular weight detection (~10 kDa for SCGB1A1)

  • Check for absence of non-specific bands

• Isotype Controls:

  • Use matched isotype antibodies to rule out non-specific binding due to Fc receptor interactions

  • Particularly important in flow cytometry applications

• Knockout/Knockdown Verification:

  • Test antibody on SCGB1A1 knockout tissues or knockdown cell lines

  • Signal should be absent or significantly reduced

Manufacturers may provide specificity data in their technical information. For example, one supplier explicitly states "No cross-reactivity in ELISA with: Human SCGB1A1" for their mouse-reactive SCGB1A1 antibody, indicating it specifically recognizes mouse but not human forms of the protein .

What are the challenges in detecting SCGB1A1 expression in different pathological conditions?

Detection of SCGB1A1 in pathological conditions presents several unique challenges due to disease-related alterations in protein expression, localization, and modification:

• Expression Level Fluctuations:

  • SCGB1A1 expression is often dysregulated in respiratory diseases

  • In inflammatory lung conditions, expression may be significantly reduced

  • Cancer may show heterogeneous expression patterns requiring careful assessment

• Post-translational Modifications:

  • Oxidative stress in disease can modify SCGB1A1 protein structure

  • These modifications may affect antibody binding efficiency

  • Different antibody clones may vary in their ability to recognize modified forms

• Cellular Redistribution:

  • In certain pathologies, SCGB1A1 may relocalize within cells

  • This may necessitate different sample preparation techniques

  • Subcellular fractionation might be required for thorough analysis

• Tissue Remodeling Effects:

  • Fibrosis and tissue remodeling can mask epitopes

  • May require optimized antigen retrieval protocols

  • Background staining can increase in highly fibrotic areas

• Protein Degradation:

  • Proteolytic environment in inflammatory conditions

  • Degraded forms may not be recognized by some antibodies

  • Fresh samples or specialized fixation may be required

To address these challenges, researchers should consider employing multiple antibody clones recognizing different epitopes and combining immunohistochemical approaches with molecular techniques such as RNA in situ hybridization to correlate protein detection with mRNA expression levels . Careful optimization of detection protocols for each specific pathological condition is often necessary.

How can researchers overcome cross-reactivity issues with SCGB1A1 antibodies?

Cross-reactivity is a significant concern when working with SCGB1A1 antibodies due to the existence of multiple secretoglobin family members with structural similarities. Several strategies can help researchers minimize and address potential cross-reactivity issues:

• Antibody Selection Strategy:

  • Choose recombinant monoclonal antibodies with validated specificity

  • Review manufacturer data on cross-reactivity testing

  • Consider antibodies raised against unique regions of SCGB1A1

• Absorption Controls:

  • Pre-absorb antibodies with recombinant proteins of related family members

  • Test reduced signal with SCGB1A1 absorption versus maintained signal with related protein absorption

• Optimization Techniques:

  • Increase antibody dilution to reduce non-specific binding

  • Optimize blocking solutions (consider protein-free blockers)

  • Adjust incubation times and temperatures

• Washing Protocol Refinement:

  • Extend washing steps after antibody incubation

  • Use detergent concentrations appropriate for reducing non-specific binding

  • Consider high-salt washes for particularly problematic cross-reactivity

• Cross-Validation Approaches:

  • Use multiple antibodies targeting different epitopes

  • Compare results from antibodies from different manufacturers

  • Verify with non-antibody-based detection methods

Manufacturers typically provide cross-reactivity data in their technical information. For example, one recombinant monoclonal antibody is specifically noted to show "No cross-reactivity in ELISA with: Human SCGB1A1" when testing for mouse SCGB1A1, demonstrating species-specific targeting without human cross-reactivity . This type of information is valuable when selecting antibodies for multi-species studies or when working with conserved protein families.

What are the optimal fixation and antigen retrieval methods for SCGB1A1 immunohistochemistry?

The selection of appropriate fixation and antigen retrieval methods significantly impacts the detection sensitivity and specificity of SCGB1A1 in immunohistochemical applications. Based on research practices and manufacturer recommendations, the following protocols have demonstrated effectiveness:

Optimal Fixation Protocols for SCGB1A1 Detection:

Recommended Antigen Retrieval Methods:

• Heat-Induced Epitope Retrieval (HIER):

  • Citrate buffer (pH 6.0) at 95-100°C for 20 minutes

  • EDTA buffer (pH 8.0-9.0) for cases with weak signal using citrate

  • Pressure cooker methods (2-3 minutes at pressure) often yield superior results

• Enzymatic Antigen Retrieval:

  • Proteinase K digestion (10-20 μg/mL for 10-15 minutes at room temperature)

  • Less commonly used but may be beneficial for heavily fixed samples

The effectiveness of these methods may vary based on specific tissue type, fixation duration, and the particular SCGB1A1 antibody clone being used. Based on immunohistochemistry images from manufacturer data, successful staining of mouse lung tissue has been achieved using paraffin-embedded sections with appropriate antigen retrieval methods .

A systematic comparison of multiple retrieval methods on identically fixed tissues is recommended when establishing a new SCGB1A1 immunostaining protocol. This approach helps identify the optimal procedure for specific research requirements.

How should researchers design blocking protocols to minimize background staining when using SCGB1A1 antibodies?

Effective blocking is crucial for minimizing background staining when using SCGB1A1 antibodies, particularly in tissues with high endogenous peroxidase activity or biotin content like lung tissue. A comprehensive blocking strategy should address multiple sources of background:

Recommended Blocking Protocol Components:

• Endogenous Peroxidase Blocking:

  • 0.3-3% hydrogen peroxide in methanol for 10-30 minutes

  • Critical for lung tissue where endogenous peroxidase activity is high

• Protein Blocking Options:

  • Normal serum blocking (5-10% from species unrelated to primary and secondary antibodies)

  • Commercial protein-free blockers for sensitive applications

  • BSA-based blockers (1-5% in PBS or TBS)

• Avidin-Biotin Blocking:

  • Essential when using biotin-based detection systems

  • Sequential avidin and biotin blocking solutions (15 minutes each)

  • Particularly important in lung tissue with endogenous biotin

• Fc Receptor Blocking:

  • Add 10% serum from the secondary antibody species

  • Commercial Fc receptor blocking solutions

  • Critical for flow cytometry applications

• Non-specific Site Saturation:

  • 0.1-0.3% Triton X-100 or 0.05% Tween-20 to reduce hydrophobic interactions

  • Include in blocking buffer for balanced membrane permeabilization

Optimization Considerations:

Testing multiple blocking protocols is advisable when establishing a new SCGB1A1 staining procedure, as the optimal approach may vary depending on the specific antibody clone, tissue type, and detection system employed .

What controls should be included when performing Western blot analysis with SCGB1A1 antibodies?

Proper controls are essential for ensuring reliable and interpretable Western blot results when detecting SCGB1A1. A comprehensive control strategy should include:

Essential Controls for SCGB1A1 Western Blot Analysis:

• Positive Control:

  • Lung tissue lysate (primary source of SCGB1A1 expression)

  • Recombinant SCGB1A1 protein standard

  • Cell lines known to express SCGB1A1 (e.g., certain lung epithelial lines)

• Negative Control:

  • Tissues or cell lines lacking SCGB1A1 expression

  • SCGB1A1 knockout or knockdown samples when available

  • Non-epithelial tissue samples with minimal SCGB1A1 expression

• Loading Control:

  • Housekeeping proteins (β-actin, GAPDH, tubulin)

  • Total protein stains (Ponceau S, SYPRO Ruby, stain-free technology)

  • Especially important due to SCGB1A1's small size (~10 kDa)

• Molecular Weight Marker:

  • Low molecular weight marker inclusive of 10 kDa range

  • Pre-stained markers for transfer efficiency monitoring

• Antibody Controls:

  • Primary antibody omission control

  • Isotype control antibody

  • Pre-absorption control with immunizing peptide

Additional Validation Approaches:

Due to SCGB1A1's small size (~10 kDa), particular attention should be paid to gel percentage (15-20% recommended), transfer conditions optimized for small proteins, and appropriate molecular weight markers that clearly define the 10 kDa region .

What are the best practices for storage and handling of SCGB1A1 recombinant monoclonal antibodies to maintain activity?

Storage Recommendations:

Handling Best Practices:

• Initial Processing:

  • Upon receipt, briefly centrifuge vials to collect contents at the bottom

  • Prepare working aliquots in sterile microcentrifuge tubes

  • Record lot number, receipt date, and expiration date on each aliquot

• Working Solution Preparation:

  • Use sterile buffers for dilution

  • Include carrier protein (0.1-1% BSA) in working dilutions to prevent adsorption

  • Prepare fresh working dilutions on the day of use when possible

• Contamination Prevention:

  • Use sterile pipette tips and containers

  • Filter buffers used for antibody dilution (0.2 μm)

  • Include preservatives (0.02-0.05% sodium azide) for dilutions stored >24 hours

• Temperature Management:

  • Avoid exposure to temperatures above 37°C

  • Thaw frozen aliquots on ice or at 4°C

  • Return to appropriate storage promptly after use

• Documentation Practices:

  • Maintain a usage log for each antibody

  • Record freeze-thaw cycles and observed performance

  • Note lot-to-lot variations in working concentration

Some SCGB1A1 antibodies are formulated as "azide and BSA free" , which may require specific handling considerations. These formulations may be preferred for certain applications (e.g., in vivo studies, functional assays) but may necessitate stricter adherence to sterile technique and consideration of shorter shelf-life for working dilutions.

How can researchers troubleshoot inconsistent results when using SCGB1A1 antibodies across different experimental platforms?

Inconsistent results across different experimental platforms when using SCGB1A1 antibodies can stem from multiple technical and biological factors. A systematic troubleshooting approach should consider platform-specific variables along with antibody characteristics:

Common Issues and Troubleshooting Strategies:

• Discrepancies Between IHC and Western Blot Results:

  • Issue: Positive IHC but negative Western blot

  • Potential causes: Conformation-dependent epitopes, protein denaturation

  • Solution: Try different antibody clones, modify extraction buffers, use non-reducing conditions

• Flow Cytometry vs. Immunofluorescence Inconsistencies:

  • Issue: Different staining patterns or intensities

  • Potential causes: Cell permeabilization differences, fixation effects

  • Solution: Standardize fixation protocols, optimize permeabilization, adjust antibody concentration

• ELISA Sensitivity Variations:

  • Issue: Inconsistent detection thresholds across platforms

  • Potential causes: Different binding conditions, buffer effects

  • Solution: Use identical recombinant standards, standardize coating conditions

Systematic Troubleshooting Approach:

Platform-Specific Considerations for SCGB1A1:

• Western Blot:

  • Due to SCGB1A1's small size (~10 kDa), use high percentage gels (15-20%)

  • Optimize transfer conditions for small proteins

  • Consider using PVDF membranes for better retention of small proteins

• Immunohistochemistry:

  • Optimize antigen retrieval methods specifically for lung tissue

  • Address lung-specific background issues (endogenous peroxidase, biotin)

  • Use amplification systems for tissues with lower expression

• Flow Cytometry:

  • Test different permeabilization methods (saponin vs. Triton X-100)

  • Optimize fixation to preserve epitopes

  • Use appropriate compensation controls for multi-color experiments

When possible, validate findings using orthogonal methods that don't rely on antibody recognition, such as mass spectrometry or transcript analysis. Consider also testing multiple antibody clones targeting different epitopes on the SCGB1A1 protein to distinguish between technical issues and biological variability .

What emerging research areas are utilizing SCGB1A1 recombinant monoclonal antibodies?

SCGB1A1 recombinant monoclonal antibodies are becoming increasingly important tools in several cutting-edge research areas at the intersection of respiratory biology, immunology, and precision medicine. These emerging fields are leveraging the high specificity and reproducibility of recombinant monoclonal antibodies to advance understanding of SCGB1A1's diverse biological roles.

Key emerging research areas include:

• Respiratory Disease Biomarker Development:

  • Using SCGB1A1 detection as prognostic/diagnostic markers in COPD, asthma, and IPF

  • Quantifying SCGB1A1 in bronchoalveolar lavage fluid and serum as disease indicators

  • Correlating SCGB1A1 expression patterns with disease progression and therapeutic response

• Lung Regeneration and Stem Cell Research:

  • Tracking Clara cell-derived progenitor populations during lung repair

  • Using SCGB1A1 as a marker for successful differentiation of stem cells into lung epithelial lineages

  • Studying the role of SCGB1A1-expressing cells in lung regeneration after injury

• COVID-19 Research Applications:

  • Investigating SCGB1A1 expression changes in SARS-CoV-2 infected lung tissues

  • Exploring potential protective roles of SCGB1A1 against viral-induced inflammation

  • Monitoring Clara cell damage and recovery in post-COVID lung pathology

• Cancer Immunology Investigations:

  • Examining SCGB1A1's immunomodulatory effects in the tumor microenvironment

  • Exploring correlations between SCGB1A1 expression and lung cancer subtypes and outcomes

  • Investigating potential therapeutic applications targeting SCGB1A1 pathways

• Environmental Toxicology Applications:

  • Using SCGB1A1 as a biomarker for environmental lung damage

  • Studying responses to air pollutants, cigarette smoke, and industrial exposures

  • Developing screening methodologies for respiratory toxicants

These research directions benefit from the precision and reproducibility offered by recombinant monoclonal antibodies against SCGB1A1, enabling more consistent results across laboratories and experimental platforms. The continued development of novel antibody formats with enhanced properties will likely further expand these applications in coming years .

How can researchers integrate SCGB1A1 antibody-based methods with other molecular techniques for comprehensive analysis?

Integrating SCGB1A1 antibody-based detection methods with complementary molecular techniques creates powerful research workflows that provide comprehensive insights into SCGB1A1 biology. Strategic integration of multiple methodologies allows researchers to correlate protein expression with genetic regulation, functional outcomes, and disease mechanisms.

Effective Integration Strategies:

• Multi-Omics Integration Approaches:

  • Combine antibody-based proteomics (IHC, Western blot) with transcriptomics (RNA-seq, qPCR)

  • Correlate protein localization with gene expression patterns in single-cell analysis

  • Integrate posttranslational modification data with protein expression levels

• Functional Correlation Methods:

  • Pair antibody-based detection with reporter assays for SCGB1A1 promoter activity

  • Connect protein expression patterns with functional assays of pulmonary protection

  • Correlate SCGB1A1 levels with inflammatory biomarkers in biological samples

• Spatial Biology Applications:

  • Combine SCGB1A1 immunofluorescence with in situ hybridization for mRNA detection

  • Use multiplexed antibody panels to assess SCGB1A1 in the context of cell type markers

  • Apply spatial transcriptomics with antibody validation of key findings

• Temporal Analysis Methods:

  • Implement time-course studies with antibody detection at multiple timepoints

  • Correlate dynamic changes in SCGB1A1 expression with disease progression markers

  • Use inducible genetic systems with antibody validation of protein expression

Workflow Examples for Common Research Questions: