PSD95 has a predicted band size of 95kDa, however in western blot gels it tends to run at a larger band size (~105kDa), this is thought to be due to post-translational modification (possibly palmitoylation) of PSD95 proteins.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration.
Use a concentration of 0.3 - 3 µg/ml. Detects a band of approximately 105 kDa (predicted molecular weight: 95 kDa).
Interacts with the cytoplasmic tail of NMDA receptor subunits and shaker-type potassium channels. Required for synaptic plasticity associated with NMDA receptor signaling. Overexpression or depletion of DLG4 changes the ratio of excitatory to inhibitory synapses in hippocampal neurons. May reduce the amplitude of ASIC3 acid-evoked currents by retaining the channel intracellularly. May regulate the intracellular trafficking of ADR1B.
Belongs to the MAGUK family. Contains 1 guanylate kinase-like domain. Contains 3 PDZ (DHR) domains. Contains 1 SH3 domain.
The PDZ domain 3 mediates interaction with ADR1B. The L27 domain near the N-terminus of isoform 2 is required for HGS/HRS-dependent targeting to postsynaptic density.
Palmitoylation of isoform 1 is required for targeting to postsynaptic density.
Cell membrane. Cell junction, synapse, postsynaptic cell membrane, postsynaptic density. Cell projection, axon. Cell junction, synapse. High levels in postsynaptic density of neurons in the forebrain. Also in presynaptic region of inhibitory synapses formed by cerebellar basket cells on axon hillocks of Purkinje cells.
Immunoprecipitation - Anti-PSD95 (phospho S295) antibody (ab16495)This image is courtesy of Chris Anderson, Wellcome Trust Sanger Institute, United Kingdom
Lane 1 - Input lane, 25-50µg mouse forebrain brain lysate probed with ab16495 PSD95 (phospho S295) antibody (1µg/ml)
Lane 2 - IP lane, ~500µg mouse forebrain brain lysate probed with ab16495 PSD95 (phospho S295) antibody (1µg/ml)
The IP lane (2) shows an enrichment of PSD95 (phospho S295) at the expected band size (~105kDa).
Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-PSD95 (phospho S295) antibody (ab16495)This image is courtesy of an abreview submitted by Sophie Pezet, CNRS, Paris, France
Immunohistochemistical detection of PSD95 (phospho S295) using antibody ab16495 on PFA perfusion-fixed mouse brain sections (30 microns). Primary antibody ab16495 was diluted at 1/300 and incubated for 18 hours @ 20°C in PBS + 0.3 % Triton X100. Secondary antibody: goat anti-rabbit Alexa Fluor 488 (1/1000). The image show immunostaining obtained in the mouse cerebellum using this antibody using direct fluorescence. The antibody stained granular cerebellar cells. Image [A] shows the staining obtained using the antibody, whereas image [B] is the staining captured with the same setup of acquisition for the negative control (lack of primary antibody). The arrows show the location of purkinje cells.