Protein fingerprints of cultured CA3-CA1 hippocampal neurons: comparative analysis of the distribution of synaptosomal and cytosolic proteins

Additional file 1:

Synaptic protein enrichment assayed by Western blots analysis. Synaptic protein enrichment assayed by Western blots analysis of obtained loading total homogenate (Tot) and purified synaptosomes (Syn) from CA3-CA1 hippocampal cultures (50 μg/lane), same method and source material used for synaptic 2D gels. Proteins were resolved on a 10% acrylamide SDS-PAGE and electrotransferred to nitrocellulose for immunolabelling with specific antibodies against well known synaptic markers Antibodies used: Anti glutamate receptors GluR1 (rabbit policlonal made in house, 1:500), P38 (Synaptic System, 1:500) synaptotagmin-1 (rabbit policlonal against the lumenal domain, made in house, 1:1000), VAMP-2 (Synaptic system,1:500), Beta-tubulin (Sigma, 1:1000). Antibodies reactivity was revealed by using appropriate HRP-conjugated secondary antibodies followed by chemiluminescence reaction and film exposure.

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Additional file 2:

List of differentially expressed proteins in the synaptosomal vs. crude cytosolic fractions.

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Additional file 3:

Statistical analysis of proteins present with different expression level in synaptosomal and crude cytosolic fractions.

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