A Sensitive PSD-95 Antibody for Synaptic Analysis

Physiology and Biophysics
State University of New York at Buffalo
Postdoctoral Researcher

Overall

Quality of Results

Ease-of-Optimization

What do these ratings mean?
Write a Review

Company:

Sigma-Aldrich

Product Name:

Monoclonal Anti-PSD95 antibody produced in mouse

Catalog Number:

P246

Clone Number:

7E3-1B8, monoclonal

We utilize the PSD-95 antibody to evaluate the expression of the postsynaptic marker of glutamatergic synapses in the prefrontal cortex. We perform a single-cell, Imaris-based analysis of the spatial density of the postsynaptic marker of excitatory synapses and correlate it with mice's cognitive performance following different manipulations.

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

Mouse samples; 40-μm thick coronal brain sections

Primary Incubation

1:1000; 4°C overnight

Blocking Agent

3% normal goat serum and 0.3% Triton X-100 in PBS for 1 h at room temperature

Secondary Incubation

1:1000; Alexa 633-labeled anti-mouse fluorescent secondary antibody

Tertiary Incubation

N/A

Detection

Leica SP8 confocal microscope with PMT and HyD detectors

Results Summary

PSD-95 plays a crucial role in the stabilization of excitatory synapses. We measure PSD-95 together with VGLUT1 immunoreactivity to evaluate glutamatergic synaptic puncta. We found a significantly reduced density of VGLUT1⁺/PSD-95⁺ puncta in mice with deficient energy metabolism in astrocytes. Mice with altered energy metabolism in astrocytes and altered expression of postsynaptic markers of glutamatergic synapses within the territories of energy-deficient astrocytes showed cognitive deficits. Images: representative confocal images of brain samples (prefrontal cortex) from different mice.

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

Delivers sensitive, robust, and reproducible results.

The Bad

Rather expensive for 100 µL.

The Bottom Line

Highly recommend, as it works as expected — it does not require any exhausting calibrations. The signal is very robust.

Share your experience with other scientists. Write a Review! »

Join the discussion