Reliable Detection of Postsynaptic Puncta in Mouse Brain Tissue

July 07, 2026

BlueskyReddit
SUNY UB
Research Scientist

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Company:

MilliporeSigma

Product Name:

Anti-PSD95 Antibody

Catalog Number:

P246

Clone Number:

Info:Supplier Page

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We used this antibody to visualize and quantify excitatory postsynaptic puncta in mouse prefrontal cortex. The antibody produced clear and well-defined punctate staining that was suitable for high-resolution confocal imaging and quantitative 3D analysis. We used PSD95 together with VGLUT1 to identify closely associated pre- and postsynaptic puncta within the territories of individual GFP-labeled astrocytes.

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

40-μm free-floating mouse prefrontal cortex brain sections

Primary Incubation

1:1000 overnight at 4°C

Blocking Agent

3% normal goat serum + 0.3% Triton X-100 in PBS, 1 hour at room temperature

Secondary Incubation

Corresponding Alexa Fluor–conjugated secondary antibody, 1:1000, 2 hours at room temperature

Tertiary Incubation

NA

Detection

Confocal microscopy (Leica SP8) and 3D image analysis using Imaris

Results Summary

The antibody generated distinct PSD95-positive puncta with sufficient contrast for automated and semiautomated 3D image analysis. In combination with VGLUT1 staining, it enabled quantitative analysis of excitatory synaptic puncta density and their spatial relationship to individual astrocyte surfaces.

DOI or PMID #

DOI: 10.1126/sciadv.adt2527

Additional Notes

Image Gallery

Summary

The Good

Clear punctate staining, good signal-to-background ratio, and excellent compatibility with quantitative 3D synapse analysis.

The Bad

Accurate puncta quantification requires consistent confocal acquisition and analysis parameters across samples.

The Bottom Line

A reliable antibody for high-resolution visualization and quantitative analysis of excitatory postsynaptic puncta in mouse brain tissue.

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