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Affinity purification of PSD-95-containing postsynaptic complexes by Lucia Vinade; Michael Chang; Michelle L. Schlief; Jennifer D. Petersen; Thomas S. Reese; Jung-Hwa Tao-Cheng; Ayse Dosemeci is a Neuroscience article available to read on EtoBox.

What is Affinity purification of PSD-95-containing postsynaptic complexes about?

## Abstract A widely used method for the preparation of postsynaptic density (PSD) fractions consists of treatment of synaptosomal membranes with Triton X‐100 and further purification by density gradient centrifugation. In the present study, the purity of this preparation was assessed by electron microscopic analysis. Thin‐section and rotary shadow immuno‐electron microscopy of the Triton X‐100‐derived PSD fraction shows many PSD‐95‐positive structures that resemble __in situ__ PSDs in shape and size. However, the fraction also includes contaminants such as CaMKII clusters, spectrin filaments and neurofilaments. We used magnetic beads coated with an antibody against PSD‐95 to further purify PSD‐95‐containing complexes from the Triton‐derived PSD fraction. Biochemical analysis of the affinity‐purified material shows a substantial reduction in the astrocytic marker glial fibrillary acidic protein and electron microscopic analysis shows mostly individual PSDs attached to magnetic beads. This preparation was used to assess the association of α‐amino‐3‐hydroxy‐5‐methylisoxazole‐4‐propionate (AMPA)‐type glutamate receptors with the PSD‐95‐containing complex. AMPA receptors are demonstrat

Who reads Affinity purification of PSD-95-containing postsynaptic complexes?

Author

Lucia Vinade; Michael Chang; Michelle L. Schlief; Jennifer D. Petersen; Thomas S. Reese; Jung-Hwa Tao-Cheng; Ayse Dosemeci

Publisher

John Wiley and Sons; Wiley (Blackwell Publishing); Blackwell Publishing Inc.; Wiley (ISSN 0022-3042)

Published

2003

Language

EN

Field

Neuroscience (Life Sciences)