PSEN2 Antibody, Biotin conjugated

What is PSEN2 and why is it a significant research target?

PSEN2 (Presenilin-2) is a protein with 448 amino acid residues and a mass of 50.1 kDa in humans. It serves as a suspected catalytic subunit of the gamma-secretase complex, an endoprotease complex that catalyzes the intramembrane cleavage of integral membrane proteins such as Notch receptors and APP (amyloid-beta precursor protein) . The protein's subcellular localization is primarily in the endoplasmic reticulum and Golgi apparatus . PSEN2's involvement in Alzheimer's disease pathology makes it a critical target for neurodegenerative disease research, as mutations in this gene have been linked to early-onset familial Alzheimer's disease. Additionally, PSEN2 serves as a valuable cellular marker for characterizing neuronal cells in research settings .

How does biotin conjugation enhance PSEN2 antibody functionality in experimental applications?

Biotin conjugation provides significant advantages for PSEN2 antibody applications through several mechanisms. First, the strong affinity between biotin and streptavidin (Kd ≈ 10^-15 M) enables highly sensitive detection protocols using streptavidin-coupled reporter molecules like HRP for Western blotting . This conjugation allows for signal amplification in detection systems, enhancing sensitivity when working with low-abundance PSEN2 protein samples. The biotin-streptavidin system also enables versatile experimental designs including proximity-dependent biotinylation assays that can reveal novel PSEN2 protein interactions . Additionally, biotin conjugation permits multiplexed experimental approaches where several target proteins can be simultaneously detected and analyzed using different detection systems.

What are the optimal storage conditions to maintain PSEN2 Antibody, Biotin conjugated activity?

To maintain optimal activity of PSEN2 Antibody, Biotin conjugated, specific storage protocols should be followed. Upon receipt, the antibody should be stored at -20°C or -80°C . For long-term storage, it is recommended to aliquot the antibody to minimize repeated freeze-thaw cycles which can degrade the protein structure and reduce functionality . When handling the biotin-conjugated antibody, exposure to light should be minimized as photodegradation can affect the biotin moiety and compromise downstream applications . The antibody is typically supplied in a buffer containing 50% glycerol with 0.03% Proclin 300 as a preservative in 0.01M PBS at pH 7.4, which helps maintain stability during storage . For working solutions, maintain the antibody on ice during experimental procedures and return to appropriate storage conditions immediately after use.

What experimental controls are essential when using PSEN2 Antibody, Biotin conjugated?

When designing experiments with PSEN2 Antibody, Biotin conjugated, several critical controls should be incorporated:

• Negative controls: Include samples without primary antibody treatment but with streptavidin detection reagents to assess non-specific binding of the detection system.

• Isotype controls: Use biotin-conjugated rabbit IgG (matching the PSEN2 antibody's isotype) to evaluate non-specific binding through the Fc region .

• Blocking controls: Implement pre-adsorption tests using the immunogen (Recombinant Human Presenilin-2 protein, 7-77AA) to confirm antibody specificity .

• Positive controls: Include samples known to express PSEN2 (such as neuronal cell lines) to validate detection systems.

• Cross-reactivity controls: While the antibody is specifically reactive to human PSEN2, testing with other species' samples can confirm specificity when working with mixed-species systems .

• Endogenous biotin controls: In biotin-rich tissues (like brain tissue), streptavidin-only controls help distinguish between endogenous biotin and antibody-conjugated biotin signals.

These controls ensure experimental rigor and facilitate accurate interpretation of results when investigating PSEN2 expression or interactions.

How can PSEN2 Antibody, Biotin conjugated be effectively utilized in proximity proteome analysis?

PSEN2 Antibody, Biotin conjugated can be strategically employed in proximity proteome analysis through the following methodological approach:

• Experimental setup: Utilize the biotin-conjugated PSEN2 antibody in conjunction with proximity labeling techniques such as BioID or TurboID to identify proteins physically proximal to PSEN2 in living cells .

• Sample preparation: Follow the protocol where biotin-labeled proteins are enriched using streptavidin pull-down methods after proximity labeling reactions .

• Analysis workflow: Perform on-bead trypsin digestion of pulled-down proteins, which allows direct mass spectrometry analysis of peptides harboring biotin-conjugated lysine (K+226) .

• Sensitivity enhancement: This approach significantly improves detection sensitivity by removing the bulk of unlabeled peptides, thereby reducing false positive results .

• Validation methodology: Confirm physical proximity of PSEN2 and identified proteins through secondary proximity-dependent biotinylation assays, similar to those used to validate interactions between TREM2 and proteins like DAP12, CISD2, and CKAP4 .

• Quantitative assessment: Implement quantitative analysis of biotin incorporation, as proximity labeling-mediated biotin incorporation has been shown to be quantifiable, allowing for strength-of-interaction assessments .

This comprehensive approach leverages the biotin conjugation to reveal novel protein interaction partners of PSEN2, potentially uncovering new insights into its role in cellular functions and disease mechanisms.

What are the optimal dilution parameters for PSEN2 Antibody, Biotin conjugated in various applications?

Determining optimal dilution parameters for PSEN2 Antibody, Biotin conjugated requires systematic titration based on the specific application:

• For ELISA applications (the primary tested application for this antibody):

  • Starting dilution range: 1:1000 to 1:5000

  • Optimization process: Perform checkerboard titration with both antigen and antibody dilutions

  • Signal-to-noise evaluation: Calculate signal-to-background ratios at each dilution to determine optimal concentration

• For potential Western blotting applications:

  • Initial recommendation: 1:200 to 1:1000 dilution

  • Blocking consideration: Use biotin-free blocking reagents to prevent interference

  • Detection system: Optimize streptavidin-HRP concentration in parallel with antibody dilution

• For immunocytochemistry/immunohistochemistry adaptations:

  • Starting dilution: 1:100 to 1:500

  • Incubation parameters: Optimize both time (2-24 hours) and temperature (4°C vs. room temperature)

  • Signal amplification: Consider tyramide signal amplification systems for enhanced sensitivity

Each laboratory should empirically determine optimal dilutions as performance may vary depending on sample type, detection system, and experimental conditions . Document optimization experiments systematically to establish reproducible protocols for specific research applications.

How can PSEN2 Antibody, Biotin conjugated be utilized in multiplex immunoassay systems?

Implementing PSEN2 Antibody, Biotin conjugated in multiplex immunoassay systems requires strategic methodological design:

• Platform selection: Incorporate the antibody into bead-based multiplex systems (e.g., Luminex) or planar array platforms where biotin-streptavidin interactions can be leveraged for immobilization or detection.

• Signal discrimination strategy: When multiplexing with other biotin-conjugated antibodies, use spectrally distinct streptavidin conjugates (different fluorophores) or spatial separation techniques.

• Sequential detection approach: Implement a sequential detection protocol where biotin-conjugated PSEN2 antibody is detected first, followed by blocking of remaining biotin binding sites before introducing additional biotin-conjugated detection reagents.

• Cross-reactivity mitigation: Pre-adsorb the PSEN2 antibody with potentially cross-reactive antigens to increase specificity, particularly important when targeting multiple antigens simultaneously.

• Dual recognition systems: Design sandwich assays where PSEN2 is captured by one antibody and detected by the biotin-conjugated PSEN2 antibody, enhancing specificity in complex samples.

• Calibration methodology: Develop internal calibration curves using recombinant PSEN2 protein standards to ensure quantitative accuracy across multiplexed targets.

This strategic approach enables researchers to simultaneously investigate PSEN2 alongside other proteins of interest, particularly valuable when studying complex signaling networks or disease mechanisms involving multiple interacting partners.

What methodological approaches can resolve contradictory results when using PSEN2 Antibody, Biotin conjugated?

When faced with contradictory results using PSEN2 Antibody, Biotin conjugated, researchers should employ the following systematic troubleshooting methodology:

• Antibody validation reassessment:

  • Verify antibody performance using positive control samples with known PSEN2 expression

  • Conduct Western blot analysis to confirm target specificity at the expected molecular weight (50.1 kDa)

  • Perform pre-adsorption tests with immunogen peptide (7-77AA of human PSEN2) to confirm specificity

• Technical parameter evaluation:

  • Systematically vary antibody concentrations across a wider range than initially tested

  • Modify incubation conditions (time, temperature, buffer composition) to optimize signal-to-noise ratio

  • Compare different detection systems (HRP vs. fluorescent streptavidin conjugates)

• Sample preparation analysis:

  • Examine how different protein extraction methods affect epitope accessibility

  • Test multiple fixation protocols if working with tissue samples

  • Consider the impact of post-translational modifications on epitope recognition

• Cross-laboratory validation:

  • Implement identical protocols across different laboratory settings to identify environment-specific variables

  • Exchange samples between laboratories to distinguish sample-specific from protocol-specific issues

• Alternative detection strategy:

  • Compare results with non-conjugated PSEN2 antibodies

  • Utilize alternative antibodies targeting different epitopes of PSEN2

  • Implement orthogonal detection methods (e.g., mass spectrometry) to verify protein identity

• Data integration approach:

  • Develop a weighted scoring system to integrate multiple experimental replicates

  • Conduct statistical meta-analysis of replicate experiments to identify outliers and consistent trends

This comprehensive troubleshooting framework enables researchers to systematically identify sources of variability and resolve contradictory experimental outcomes.

How can PSEN2 Antibody, Biotin conjugated be adapted for in vivo biotinylation studies?

Adapting PSEN2 Antibody, Biotin conjugated for in vivo biotinylation studies requires specialized methodological considerations:

• Expression system development: Create a system similar to the yeast-based diploid expression system used for in vivo biotinylated recombinant antibodies, where one construct carries the PSEN2-targeting sequence and another provides biotin ligase activity .

• Secretory pathway engineering: Incorporate KEX2 golgi-localization sequences to ensure biotin ligase (BirA) reaches the appropriate cellular compartment for effective biotinylation within the secretory pathway .

• Construct design strategy: Generate a fusion construct containing:

  • PSEN2-binding domain from the validated antibody

  • IgA1 hinge region for flexibility

  • Biotin acceptor site (BCCP) at the C-terminus to accept biotin conjugation

• Biotin ligase integration: Co-express the E. coli biotin ligase (BirA) to catalyze site-specific biotinylation of the acceptor sequence in the secretory pathway .

• Selection methodology: Implement appropriate selection markers (e.g., URA3, TRP1) to maintain both constructs in the expression system .

• Validation protocol: Confirm successful biotinylation through:

  • Western blot analysis with streptavidin-HRP

  • Mass spectrometry detection of biotin-conjugated lysine residues

  • Functional binding assays to verify target recognition is maintained

This adaptation leverages principles from established in vivo biotinylation systems to create PSEN2-specific biobodies that offer advantages for tracking PSEN2 interactions in living cells with minimal disruption to native protein environments.

How does polyclonal PSEN2 Antibody, Biotin conjugated compare with monoclonal alternatives in specific research applications?

The selection between polyclonal PSEN2 Antibody, Biotin conjugated and monoclonal alternatives should be based on application-specific considerations:

The polyclonal PSEN2 Antibody, Biotin conjugated (derived from rabbit hosts) offers advantages in detection sensitivity and robustness against epitope masking or protein degradation . Conversely, monoclonal alternatives provide superior reproducibility and specificity. For initial characterization studies or applications where signal amplification is critical, the polyclonal version represented in the search results provides adequate performance, particularly in ELISA applications .

What criteria should guide the selection between different applications of PSEN2 Antibody, Biotin conjugated?

When determining the optimal application for PSEN2 Antibody, Biotin conjugated, researchers should consider these methodological selection criteria:

• Primary validated application: The antibody has been specifically validated for ELISA applications , making this the most reliable starting point for research applications.

• Sample type compatibility matrix:

  Sample Type	Application Suitability	Optimization Requirements
  Cell lysates	High (for ELISA)	Requires appropriate extraction buffers
  Tissue extracts	Moderate	May need additional blocking steps
  Fixed tissues	Limited	Not primarily validated for IHC
  Purified proteins	Very high	Optimal for direct detection

• Detection limit considerations: For studies requiring detection of low abundance PSEN2, implement signal amplification systems compatible with biotin-streptavidin interactions.

• Specificity requirements analysis: For applications requiring distinction between PSEN2 isoforms or closely related proteins (like PSEN1), additional validation steps should be performed.

• Buffer compatibility assessment: The antibody formulation (50% Glycerol, 0.01M PBS, pH 7.4, 0.03% Proclin 300) may impact compatibility with certain application buffers - dilution strategies should account for this.

• Multiplexing potential evaluation: The biotin conjugation facilitates integration into multiplex detection systems but requires careful design to avoid signal interference.

The antibody's reactivity to human PSEN2 (particularly the 7-77AA region) makes it most suitable for studies focused on human samples or models expressing human PSEN2 variants. For applications beyond ELISA, researchers should conduct preliminary validation studies to verify performance in their specific experimental context.

What methodological approaches can enhance detection of PSEN2 protein-protein interactions using biotin-conjugated antibodies?

To optimize detection of PSEN2 protein-protein interactions using biotin-conjugated antibodies, implement these advanced methodological approaches:

• Proximity-dependent biotinylation approach: Adapt the super-resolution proximity labeling technique described in the literature for TREM2 interactome studies :

  • Express PSEN2 fused to a biotin ligase (TurboID) in microglial or neuronal cell models

  • Perform in situ biotinylation to label proteins in proximity to PSEN2

  • Use the biotin-conjugated PSEN2 antibody for secondary confirmation of interactions

• Streptavidin pull-down optimization:

  • Implement on-bead trypsin digestion protocols to enhance detection sensitivity

  • Analyze peptides harboring biotin-conjugated lysine (K+226) by mass spectrometry

  • This approach significantly reduces false positives by removing unlabeled peptides

• Interaction validation methodology:

  • Perform reciprocal co-immunoprecipitation with antibodies against candidate interactors

  • Conduct proximity-dependent biotinylation assays with expression constructs for each potential interactor

  • Analyze biotin incorporation levels quantitatively to assess interaction strength

• Subcellular localization refinement:

  • Focus investigation on ER and Golgi compartments where PSEN2 is predominantly localized

  • Use appropriate fractionation techniques to enrich for these organelles before interaction analysis

  • Incorporate appropriate compartment-specific markers as controls

• Cross-linking strategy integration:

  • Implement reversible cross-linking protocols before immunoprecipitation to capture transient interactions

  • Optimize cross-linker type and concentration for PSEN2's membrane environment

  • Include appropriate controls to distinguish specific from non-specific cross-linking

This comprehensive methodological framework significantly enhances the detection and validation of PSEN2 protein interactions, providing deeper insights into its functional networks in normal and pathological states.

How should researchers interpret post-translational modifications of PSEN2 in the context of antibody recognition?

When investigating post-translational modifications (PTMs) of PSEN2 using biotin-conjugated antibodies, researchers should employ this systematic interpretation framework:

• Epitope mapping analysis:

  • The PSEN2 Antibody, Biotin conjugated targets the 7-77AA region of human PSEN2

  • Evaluate whether known or predicted PTM sites overlap with this region

  • Determine potential impact of modifications on epitope recognition through structural modeling

• Modification-specific detection strategy:

  • Known PTMs of PSEN2 include phosphorylation and proteolytic processing

  • PSEN2 undergoes endoproteolysis to produce N-terminal (NTF) and C-terminal (CTF) fragments

  • Determine whether the 7-77AA epitope remains intact in these processed forms

• Comparative analysis methodology:

  • Implement parallel detection with phospho-specific antibodies if studying phosphorylation events

  • Compare detection patterns before and after phosphatase treatment

  • Use size separation techniques to distinguish full-length (50.1 kDa) from processed fragments

• Functional correlation approach:

  • Correlate detection efficiency with functional states of PSEN2 in the gamma-secretase complex

  • Assess antibody recognition in different cellular compartments where PTM status may vary

  • Evaluate detection in disease models where abnormal PTM patterns may occur

• Technical validation protocol:

  • Include recombinant PSEN2 with defined modification states as controls

  • Use mass spectrometry to independently verify PTM status in experimental samples

  • Implement multiple antibodies targeting different epitopes to create a comprehensive detection profile

This structured interpretation framework enables researchers to accurately assess how PTMs influence PSEN2 antibody recognition, providing critical context for data interpretation in both basic research and disease-focused studies.

How can PSEN2 Antibody, Biotin conjugated contribute to Alzheimer's disease research methodologies?

PSEN2 Antibody, Biotin conjugated offers several methodological advantages for Alzheimer's disease (AD) research applications:

• Gamma-secretase activity assessment:

  • PSEN2 functions as a suspected catalytic subunit of the gamma-secretase complex that processes APP

  • Use the antibody to immunoprecipitate active gamma-secretase complexes via biotin-streptavidin pull-down

  • Correlate PSEN2 detection with enzymatic activity measurements in patient-derived samples

• Disease-associated mutation analysis:

  • PSEN2 mutations are linked to early-onset familial AD (aliases include AD4, AD5, AD3LP)

  • Compare antibody recognition efficiency between wild-type and mutant PSEN2 forms

  • Develop detection protocols for specific pathogenic variants in patient-derived models

• Biomarker development methodology:

  • Implement the antibody in multiplexed biomarker panels targeting AD pathology

  • Leverage biotin-streptavidin systems for signal amplification in diagnostic assays

  • Correlate PSEN2 detection patterns with clinical parameters and disease progression

• Therapeutic target validation:

  • Use the antibody to track changes in PSEN2 expression or processing following experimental treatments

  • Develop competition assays to screen for compounds that modulate PSEN2 interactions

  • Monitor drug effects on PSEN2 localization and complex formation

• Cross-species comparative analysis:

  • Although primarily reactive to human PSEN2, evaluate cross-reactivity with model organism orthologs

  • Develop protocols for detecting human PSEN2 in humanized mouse models of AD

  • Compare PSEN2 expression patterns across species to identify conserved disease mechanisms

This methodological framework demonstrates how PSEN2 Antibody, Biotin conjugated can advance multiple aspects of Alzheimer's disease research, from basic pathophysiology to translational applications in diagnostics and therapeutics.

What methodological considerations are important when using PSEN2 Antibody, Biotin conjugated in cellular models of neurodegeneration?

When employing PSEN2 Antibody, Biotin conjugated in cellular models of neurodegeneration, researchers should implement these specialized methodological considerations:

• Cell model selection criteria:

  • Choose models with appropriate PSEN2 expression profiles relevant to human pathology

  • Consider iPSC-derived neurons or microglial cells that better recapitulate human neurodegenerative processes

  • Validate antibody performance in each cellular system prior to experimental applications

• Subcellular localization analysis protocol:

  • Implement co-localization studies with markers for ER and Golgi where PSEN2 is primarily located

  • Assess changes in localization patterns under disease-relevant stressors

  • Quantify distribution patterns using appropriate imaging and analytical tools

• Protein complex integrity assessment:

  • Evaluate PSEN2 incorporation into the gamma-secretase complex under normal and pathological conditions

  • Develop native PAGE techniques compatible with subsequent biotin-streptavidin detection

  • Compare complex formation efficiency between wild-type and mutant PSEN2 variants

• Signal transduction pathway analysis:

  • Investigate PSEN2's role in Notch signaling pathways using the biotin-conjugated antibody

  • Develop multiplexed detection systems for PSEN2 and other signaling components

  • Correlate PSEN2 detection with downstream signaling outputs

• Temporal dynamics monitoring:

  • Implement time-course studies to track PSEN2 expression and processing during cellular stress or differentiation

  • Develop pulse-chase protocols compatible with biotin-conjugated antibody detection

  • Correlate temporal changes in PSEN2 with disease-relevant phenotypes

• Cross-talk mechanism evaluation:

  • Investigate interactions between PSEN2 and other neurodegeneration-associated proteins

  • Adapt proximity-dependent biotinylation techniques to identify novel PSEN2 interactors in neuronal cells

  • Validate interactions using orthogonal approaches like co-immunoprecipitation

This comprehensive methodological framework enables researchers to maximize the utility of PSEN2 Antibody, Biotin conjugated in cellular models of neurodegeneration, advancing understanding of PSEN2's role in disease mechanisms.