DLGAP2 Antibody

What is DLGAP2 and why is it significant in neuroscience research?

DLGAP2, also known as SAP90/PSD-95-associated protein 2 (SAPAP2), is a postsynaptic scaffolding protein that plays a crucial role in the molecular organization of synapses and neuronal cell signaling. It functions as an adapter protein linking ion channels to the subsynaptic cytoskeleton and may induce enrichment of PSD-95/SAP90 at the plasma membrane . DLGAP2 has gained significant research interest due to its associations with autism spectrum disorders, schizophrenia, and more recently, with age-related cognitive decline and Alzheimer's dementia . This makes DLGAP2 antibodies essential tools for investigating the molecular basis of these neurological conditions and potential therapeutic targets.

What applications are DLGAP2 antibodies suitable for?

DLGAP2 antibodies are compatible with multiple experimental applications, providing researchers flexibility in their methodological approaches. Commercial antibodies have been validated for:

• Western Blotting (WB)

• Immunohistochemistry with paraffin-embedded sections (IHC-P)

• Immunohistochemistry with frozen sections (IHC-fro)

• Immunocytochemistry/Immunofluorescence (ICC/IF)

• ELISA assays

These applications allow researchers to detect and quantify DLGAP2 in various experimental contexts, from tissue sections to cell cultures and protein lysates . When selecting an antibody, it's important to verify that it has been validated for your specific application and species of interest.

What species reactivity is available for DLGAP2 antibodies?

Currently available DLGAP2 antibodies demonstrate reactivity primarily with human and rat samples, with predicted reactivity for mouse samples based on sequence homology . When planning experiments, it's crucial to select antibodies specifically validated for your species of interest. Cross-reactivity between species often depends on the specific epitope targeted by the antibody, which is typically a synthetic peptide corresponding to a region within the DLGAP2 protein.

How should DLGAP2 antibodies be stored and handled for optimal performance?

For optimal antibody performance, DLGAP2 antibodies should be stored according to manufacturer specifications, typically at -20°C for long-term storage and at 4°C for short-term use. Avoid repeated freeze-thaw cycles by aliquoting the antibody upon first thawing. When working with the antibody, maintain cold chain protocols and use appropriate buffers as recommended in the product documentation. Proper storage and handling are essential for maintaining antibody specificity and sensitivity in experimental applications.

What controls should be included when validating DLGAP2 antibody specificity?

When validating DLGAP2 antibody specificity, a comprehensive approach should include multiple controls:

• Positive controls: Use tissues or cell lines known to express DLGAP2 (such as brain tissue or neuronal cell lines)

• Negative controls: Include samples from DLGAP2 knockout models or use siRNA knockdown cells

• Peptide competition assays: Pre-incubate the antibody with the immunizing peptide to confirm binding specificity

• Multiple antibody validation: Use antibodies targeting different epitopes of DLGAP2 to confirm consistent localization patterns

• Cross-reactivity assessment: Test against closely related proteins (e.g., other DLGAP family members)

Additionally, researchers should be aware that the predicted molecular weight of DLGAP2 is approximately 117 kDa , which serves as a reference point when evaluating Western blot results.

How can DLGAP2 expression be effectively measured in relation to cognitive decline studies?

Recent research has established associations between DLGAP2 expression and cognitive decline, particularly in the context of Alzheimer's disease . When designing experiments to investigate these relationships, researchers should consider:

• Tissue selection: Focus on brain regions relevant to cognitive function, particularly the dorsolateral prefrontal cortex (DLPFC), which has shown significant associations between DLGAP2 expression and cognitive outcomes

• Multiple measurement methodologies:

  • RNA expression analysis via qPCR or RNA sequencing

  • Protein quantification via Western blotting

  • Mass spectrometry for precise protein quantification (tandem mass tag mass spectrometry has been successfully employed)

  • Immunohistochemistry for spatial distribution analysis

• Normalization strategies: When studying neurodegenerative conditions, account for neuronal loss by:

  • Including neuronal markers like ENO2

  • Using laser-capture microdissection to isolate specific neuronal populations

  • Normalizing to housekeeping genes stable in neurodegeneration

Studies have demonstrated that lower levels of DLGAP2 mRNA in the DLPFC are associated with poorer cognitive performance and faster cognitive decline, particularly in individuals with clinically diagnosed Alzheimer's disease .

What are the key considerations when using DLGAP2 antibodies to investigate neuropathological correlations?

When investigating correlations between DLGAP2 and neuropathological features, researchers should consider:

• Co-localization studies: Design experiments to assess DLGAP2 localization relative to pathological hallmarks like amyloid plaques and neurofibrillary tangles

• Quantitative approaches: Utilize image analysis software to quantify:

  • DLGAP2 immunoreactivity

  • Proximity to pathological structures

  • Co-localization coefficients

• Temporal dynamics: Consider the temporal relationship between DLGAP2 expression changes and the development of pathology through longitudinal studies or staged tissue samples

• Causality assessment: Design experiments to determine whether DLGAP2 alteration is causative or consequential to pathology development

Research has established that lower levels of DLGAP2 are associated with greater β-amyloid load and more neurofibrillary tangles in the DLPFC, suggesting a potential relationship with Alzheimer's disease pathogenesis .

How should antibody dilution optimization be approached for DLGAP2 detection?

Optimizing antibody dilutions is crucial for achieving specific signal with minimal background. For DLGAP2 antibodies:

• Western blotting: Begin with manufacturer-recommended dilutions (typically 0.5-1 μg/mL) and perform a dilution series to determine optimal concentration

• Immunohistochemistry: Start with recommended dilutions (often around 5 μg/mL for DLGAP2) and adjust based on:

  • Signal intensity

  • Background levels

  • Specific tissue processing methods

• Immunofluorescence: Initial concentrations around 20 μg/mL may be appropriate , with optimization considering:

  • Fixation method

  • Permeabilization protocol

  • Blocking reagents

  • Secondary antibody selection

Document all optimization steps systematically, as dilution requirements may vary between tissue types and experimental conditions.

What challenges might arise when detecting DLGAP2 in neural tissues and how can they be addressed?

Detecting DLGAP2 in neural tissues presents several challenges:

• High background in neural tissue:

  • Solution: Use more stringent blocking with 5-10% normal serum from the species of the secondary antibody

  • Implement longer blocking times (2+ hours)

  • Consider adding 0.1-0.3% Triton X-100 for better penetration

• Limited epitope accessibility:

  • Solution: Optimize antigen retrieval methods (heat-induced epitope retrieval with citrate buffer or enzymatic retrieval)

  • Test multiple fixation protocols to determine optimal epitope preservation

• Signal specificity concerns:

  • Solution: Include peptide competition controls

  • Use tissues from DLGAP2 knockout models as negative controls

  • Compare staining patterns across multiple antibodies targeting different DLGAP2 epitopes

• Variable expression levels:

  • Solution: Consider signal amplification methods (TSA amplification, polymer-based detection systems)

  • Optimize exposure settings in imaging

How can DLGAP2 antibodies be effectively used to study dendritic spine morphology?

DLGAP2 is critical for dendritic spine structure and function, with research showing associations between DLGAP2 expression, spine morphology, and cognitive outcomes . When designing experiments to investigate these relationships:

• Co-labeling strategies:

  • Combine DLGAP2 antibodies with other postsynaptic markers (PSD-95) and presynaptic markers (synaptophysin)

  • Include cytoskeletal markers (F-actin) for spine morphology assessment

• High-resolution imaging approaches:

  • Use super-resolution microscopy (STED, STORM, SIM) for detailed spine morphology

  • Confocal microscopy with deconvolution for co-localization studies

  • Consider electron microscopy with immunogold labeling for ultrastructural analysis

• Quantitative analysis:

  • Measure spine density, head diameter, neck length, and shape classifications

  • Quantify DLGAP2 enrichment within different spine types

  • Correlate DLGAP2 levels with spine maturation states

• Experimental models:

  • Compare wild-type with DLGAP2 mutant models to assess spine phenotypes

  • Use time-lapse imaging to track dynamic changes in DLGAP2 distribution during spine remodeling

How are DLGAP2 antibodies used to investigate autism spectrum disorders and schizophrenia?

DLGAP2 antibodies serve as valuable tools for investigating the molecular basis of autism spectrum disorders and schizophrenia, conditions that have been associated with DLGAP2 abnormalities . Research approaches include:

• Postmortem tissue analysis:

  • Compare DLGAP2 expression and localization in brain regions implicated in these disorders

  • Correlate with genetic variants identified in patient populations

  • Assess relationships with other synaptic proteins known to be dysregulated in these conditions

• Cellular models:

  • Use patient-derived iPSCs differentiated into neurons to examine DLGAP2 expression patterns

  • Investigate activity-dependent changes in DLGAP2 localization

  • Study the impact of disease-associated DLGAP2 variants on protein function

• Animal models:

  • Utilize DLGAP2 knockout or mutant models to characterize behavioral phenotypes

  • Correlate behavioral changes with alterations in synaptic architecture

  • Test interventions targeting DLGAP2-associated pathways

These approaches help elucidate how DLGAP2 dysfunction may contribute to synaptic abnormalities associated with neurodevelopmental disorders.

What is the significance of DLGAP2 in Alzheimer's disease research and how can antibodies help investigate this connection?

Recent research has identified DLGAP2 as a potential regulator of age-related cognitive decline and Alzheimer's dementia, making DLGAP2 antibodies increasingly important in this field . Key research applications include:

• Expression studies in Alzheimer's progression:

  • DLGAP2 expression is decreased in mild cognitive impairment (MCI) and Alzheimer's disease compared to normal cognition

  • This decrease occurs independent of frank neurodegeneration, suggesting specific dysregulation

• Correlation with pathological hallmarks:

  • Lower DLGAP2 levels are associated with greater β-amyloid load and more neurofibrillary tangles

  • No associations were observed with non-Alzheimer neuropathologies, suggesting specificity

• Genetic association studies:

  • SNPs within and near DLGAP2 have been associated with Alzheimer's disease in both European ancestry (rs2957061) and African American (chr8:1316870) populations

  • Methylation patterns in the DLGAP2 region have been associated with cognitive outcomes

• Experimental approaches using antibodies:

  • Evaluate DLGAP2 protein levels in different disease stages

  • Assess co-localization with pathological markers

  • Monitor changes in subcellular distribution during disease progression

  • Investigate relationships with synaptic loss patterns

How can DLGAP2 antibodies contribute to translational research between animal models and human studies?

DLGAP2 antibodies facilitate translational research connecting findings from animal models to human studies:

• Cross-species validation:

  • Use antibodies validated for multiple species to compare DLGAP2 expression patterns

  • Confirm that identified mechanisms in mouse models translate to human tissues

  • Validate findings from the Diversity Outbred mouse population, which has identified DLGAP2 as a regulator of working memory decline

• Biomarker development:

  • Investigate whether DLGAP2 protein levels in accessible biospecimens correlate with brain expression

  • Determine whether DLGAP2 alterations precede cognitive symptoms

  • Assess potential as a disease progression marker

• Therapeutic target validation:

  • Use antibodies to confirm target engagement in preclinical studies

  • Monitor DLGAP2 expression changes in response to experimental therapeutics

  • Develop assays to screen compounds that modulate DLGAP2 expression or function

• Comparative pathology:

  • Establish whether DLGAP2-associated pathology in mouse models recapitulates human disease features

  • Determine if spine morphology alterations associated with DLGAP2 variation in mice are also present in human samples

How might single-cell approaches enhance DLGAP2 antibody-based research?

Integrating DLGAP2 antibodies with single-cell technologies offers exciting opportunities:

• Single-cell protein analysis:

  • Combine DLGAP2 antibodies with mass cytometry (CyTOF) for high-dimensional protein profiling

  • Use imaging mass cytometry to maintain spatial context while analyzing multiple proteins simultaneously

  • Implement proximity ligation assays to investigate protein-protein interactions at single-cell resolution

• Spatial transcriptomics integration:

  • Correlate DLGAP2 protein localization with gene expression patterns in the same tissue section

  • Identify cell type-specific associations between DLGAP2 and disease states

  • Map regional vulnerability based on DLGAP2 expression patterns

• Methodological considerations:

  • Validate antibody specificity at single-cell resolution

  • Optimize fixation and permeabilization for multimodal analyses

  • Develop computational pipelines to integrate protein and transcriptomic data

These approaches could reveal cell type-specific roles of DLGAP2 in neurological disorders and identify new therapeutic opportunities.

What emerging techniques might enhance the specificity and sensitivity of DLGAP2 detection?

Several emerging technologies may enhance DLGAP2 detection:

• Advanced antibody engineering:

  • Single-domain antibodies (nanobodies) for improved penetration and resolution

  • Recombinant antibody fragments with enhanced epitope accessibility

  • Site-specific conjugation strategies for optimal fluorophore positioning

• Novel imaging approaches:

  • Expansion microscopy to physically enlarge specimens for improved resolution

  • Lattice light-sheet microscopy for rapid 3D imaging with reduced phototoxicity

  • MINFLUX nanoscopy for molecular-scale resolution of protein localization

• Multiplexed detection methods:

  • Sequential immunofluorescence with multiplexed antibody detection (>40 proteins)

  • DNA-barcoded antibodies for highly multiplexed protein mapping

  • Mass spectrometry imaging for label-free protein identification

• Live-cell applications:

  • Intrabodies derived from DLGAP2 antibodies for live-cell tracking

  • Split-GFP complementation systems for monitoring protein interactions

  • Optogenetic tagging strategies for dynamic visualization

These techniques would provide unprecedented insights into DLGAP2 dynamics and functions in neural circuits.