PSG6 Antibody

What is PSG6 and why is it important in reproductive research?

PSG6 (pregnancy-specific beta-1-glycoprotein 6) belongs to the pregnancy-specific glycoprotein family and is primarily produced by placental syncytiotrophoblasts. This protein plays a critical role in promoting immune tolerance during pregnancy and modulating maternal immune responses to support fetal development . High levels of PSG biosynthesis are restricted to the placenta syncytiotrophoblast and are essential for maintaining normal gestation in mammalian species . Research focused on PSG6 provides valuable insights into pregnancy-related disorders, immune-related complications in pregnancy, and potential therapeutic interventions targeting this protein .

What types of PSG6 antibodies are available for research applications?

Research-grade PSG6 antibodies are available in several formats:

• Host species and clonality:

  • Polyclonal antibodies (primarily rabbit-derived)

  • Monoclonal antibodies (mouse-derived)

• Target epitope variations:

  • Antibodies targeting N-terminal regions (AA 22-48)

  • Antibodies targeting central regions (AA 35-300)

  • Full-length protein antibodies (Met1-Gly424)

• Conjugation options:

  • Unconjugated primary antibodies

  • Conjugated versions (HRP, FITC, Biotin)

What are the common applications of PSG6 antibodies in research?

Based on the available data, PSG6 antibodies are validated for several key applications:

• Western Blot (WB): For detecting PSG6 protein expression in cell lysates, tissue extracts, and biological fluids like serum from pregnant donors

• Enzyme-Linked Immunosorbent Assay (ELISA): For quantitative measurement of PSG6 levels

• Immunohistochemistry (IHC): For visualizing PSG6 localization in tissue sections, particularly in placental syncytiotrophoblasts

• Cross-application considerations: Some antibodies are validated for multiple techniques, allowing consistent detection across different experimental approaches

How do I select the appropriate PSG6 antibody for my experiment?

When selecting a PSG6 antibody, consider these methodological factors:

• Experimental application compatibility: Ensure the antibody has been validated for your intended application (WB, ELISA, IHC). For example, MAB8598 is validated for both Western blot and IHC applications , while others may be optimized specifically for ELISA or Western blot .

• Epitope specificity needs: Different antibodies target specific regions of PSG6. Consider which epitope is relevant to your research question and whether potential modifications or protein interactions might mask this region.

• Species reactivity requirements: Most PSG6 antibodies are specific to human PSG6 . If working with animal models, verify cross-reactivity with your species of interest.

• Validation data availability: Review available performance data, such as Western blot images showing detection in pregnant vs. non-pregnant donor samples or staining patterns in placental tissue sections.

• Sample type compatibility: Consider whether your antibody has been validated in your specific sample type (cell lines, tissues, biological fluids).

What are the optimal storage conditions for PSG6 antibodies?

Based on manufacturer recommendations, optimal storage conditions include:

• Temperature requirements:

  • Long-term storage: -20°C to -70°C for 6-12 months

  • Short-term storage: 2-8°C for up to 1 month after reconstitution

• Buffer compositions:

  • 50% Glycerol with 0.01M PBS (pH 7.4) and 0.03% Proclin 300

  • PBS with 0.02% sodium azide and 50% glycerol (pH 7.3)

  • 0.1M Tris-Glycine (pH 7) with 10% Glycerol and 0.01% Thimerosal

• Handling precautions:

  • Avoid repeated freeze-thaw cycles by creating working aliquots

  • Use manual defrost freezers rather than auto-defrost models

  • Maintain sterile conditions after reconstitution for maximum stability

How can I validate the specificity of a PSG6 antibody for my experimental system?

Rigorous validation ensures reliable results when working with PSG6 antibodies:

• Biological controls for validation:

  • Use serum samples from pregnant donors as positive controls and non-pregnant donors as negative controls as demonstrated in R&D Systems' Western blot data

  • Include placental tissue (particularly syncytiotrophoblasts) as a positive control tissue

  • Consider HepG2 cell lysates as an additional positive control

• Technical validation approaches:

  • Conduct peptide competition assays using the immunogen sequence

  • Compare results across antibodies targeting different PSG6 epitopes

  • Correlate protein detection with mRNA expression data

  • Verify expected molecular weight patterns (calculated 48 kDa vs. observed ~75 kDa glycosylated form)

• Specificity considerations:

  • Account for potential cross-reactivity with other PSG family members

  • Consider post-translational modifications that may affect epitope recognition

  • Validate under both reducing and non-reducing conditions if appropriate

Western Blotting:

• Recommended dilutions: Range from 1:500-1:5000 depending on the antibody

• Sample preparation: Strong detection in serum from pregnant donors but not non-pregnant donors

• Expected band size: Approximately 75 kDa under reducing conditions (glycosylated form) versus calculated 48 kDa

• Membrane type: PVDF membrane recommended

• Detection systems: HRP-conjugated secondary antibodies work well with standard ECL systems

Immunohistochemistry:

• Recommended concentration: 5 μg/mL for paraffin-embedded sections

• Incubation conditions: 1 hour at room temperature

• Detection systems: Anti-Mouse IgG VisUCyte HRP Polymer Antibody with DAB visualization

• Counterstaining: Hematoxylin provides good nuclear contrast

• Expected localization: Specific staining in syncytiotrophoblasts of placental tissue

ELISA:

• Recommended dilutions: 1:2000-1:10000

• Sample types: Validated for human samples

• Optimization approaches: Titration series to determine optimal concentration

How do I troubleshoot inconsistent results with PSG6 antibodies in Western blot applications?

When troubleshooting Western blot issues with PSG6 antibodies:

• Sample-related considerations:

  • Ensure samples contain PSG6 (pregnant donor serum or placental tissue extracts are reliable positive controls)

  • Add protease inhibitors during sample preparation to prevent degradation

  • Consider glycoprotein-specific extraction methods for improved yield

• Technical optimization:

  • Adjust antibody concentration (try a range from 0.5-6 μg/mL)

  • Modify blocking conditions (BSA vs. milk-based blockers)

  • Extend primary antibody incubation time (overnight at 4°C)

  • Optimize transfer conditions for high molecular weight glycoproteins

• Detection system troubleshooting:

  • Verify secondary antibody compatibility and activity

  • Try more sensitive ECL substrates for low abundance detection

  • Adjust exposure times to capture optimal signal

• Molecular weight considerations:

  • Expected band at approximately 75 kDa (glycosylated form) rather than the calculated 48 kDa

  • Multiple bands may represent different glycosylation states

  • Compare reducing vs. non-reducing conditions

What are the considerations for studying PSG6 expression patterns during different stages of pregnancy?

For studying temporal PSG6 expression:

• Sample collection strategy:

  • Establish consistent gestational age timeline for sampling

  • Consider both maternal (serum) and placental samples

  • Include pre-pregnancy, early, middle, and late gestational samples

• Quantitative approaches:

  • Pair Western blot analysis with densitometry for semi-quantitative assessment

  • Develop quantitative ELISA protocols with recombinant PSG6 standards

  • Use digital image analysis for quantifying IHC staining intensity in tissue sections

• Expression pattern analysis:

  • Correlate with syncytiotrophoblast development stages

  • Compare with other PSG family members' expression patterns

  • Relate to pregnancy hormones and immune parameters

• Technical considerations:

  • Maintain consistent protocols across all gestational time points

  • Include appropriate housekeeping controls for normalization

  • Account for increasing blood volume during pregnancy when analyzing serum levels

How can PSG6 antibodies be used to investigate the role of PSG6 in immune modulation during pregnancy?

For investigating PSG6's immunomodulatory functions:

• Co-culture experimental designs:

  • Isolate primary immune cells and treat with recombinant PSG6

  • Block PSG6 function in placental explant cultures using antibodies

  • Analyze cytokine production profiles before and after PSG6 exposure

• Mechanistic studies:

  • Use PSG6 antibodies to immunoprecipitate potential binding partners

  • Block PSG6-receptor interactions using antibodies

  • Investigate intracellular signaling pathway activation following PSG6 treatment

• Tissue-specific analyses:

  • Perform dual immunofluorescence staining to identify PSG6-producing cells and interacting immune cells

  • Microdissect specific regions of the maternal-fetal interface for protein expression analysis

  • Compare normal vs. pathological pregnancy tissues for altered PSG6 expression

• Functional readouts:

  • Measure changes in immune cell activation markers

  • Assess proliferation and cytokine production in response to PSG6

  • Evaluate migration and adhesion properties of immune cells in the presence of PSG6

Comparison of Available PSG6 Antibodies

PSG6 Detection in Biological Samples

The following data summarizes PSG6 detection across various sample types:

• Serum analysis:

  • Present in serum from pregnant donors

  • Absent in serum from non-pregnant donors

  • Detected as a ~75 kDa band under reducing conditions

• Tissue expression:

  • Strong localization to syncytiotrophoblasts in placental tissue

  • Visualized using DAB (brown) with hematoxylin counterstain (blue)

  • Requires appropriate fixation and antigen retrieval protocols

• Cell line expression:

  • Detectable in HepG2 whole cell lysates

  • Suggests potential expression beyond placental tissues in certain contexts

Western Blot Performance:

• Clear distinction between pregnant vs. non-pregnant serum samples

• Specific band at approximately 75 kDa under reducing conditions

• Optimal antibody concentrations between 0.5-6 μg/mL depending on sample type

Immunohistochemistry Performance:

• Strong, specific staining of syncytiotrophoblasts in human placental sections

• Optimal concentration of 5 μg/mL for paraffin-embedded tissues

• Effective visualization using HRP-polymer detection systems with DAB

Emerging Applications of PSG6 Antibodies

Recent advances suggest several promising research directions:

• Biomarker development:

  • Potential use of PSG6 as a biomarker for pregnancy complications

  • Development of quantitative assays for clinical applications

  • Correlation with other established pregnancy markers

• Therapeutic target exploration:

  • Investigation of PSG6 for the development of novel interventions for pregnancy-related disorders

  • Modulation of immune responses in pregnancy complications

  • Recombinant PSG6 as a potential therapeutic agent

• Mechanistic investigations:

  • Detailed studies of PSG6-mediated signaling pathways

  • Interaction analysis with immune cell receptors

  • Structure-function relationships in immune modulation

Technical Advancements in PSG6 Research

Ongoing technical developments include:

• Antibody engineering:

  • Development of antibodies with enhanced specificity and sensitivity

  • Creation of additional conjugated formats for multiparameter analyses

  • Generation of neutralizing antibodies for functional studies

• Detection methodologies:

  • Single-cell analysis approaches

  • Multiplexed imaging systems

  • High-throughput screening platforms