Western blot - PSD95 antibody - Synaptic Marker (ab18258)

All lanes : Anti-PSD95 antibody - Synaptic Marker (ab18258) at 1 µg/ml

Lane 1 : wildtype mouse brain tissue lysate
Lane 2 : PSD95 knockout mouse brain tissue lysate

Lysates/proteins at 20 µg per lane.


Performed under reducing conditions.

Predicted band size : 80 kDa
Observed band size : ~80 kDa (why is the actual band size different from the predicted?)

Immunoprecipitation - PSD95 antibody - Synaptic Marker (ab18258)

Lane 1 - Input lane 500ug mouse brain lysate

Lane 2 - IP lane 50ug mouse brain lysate

NB: as expected, an enrichment of PSD95 protein is observed in the IP lane.

Chris Anderson, Wellcome Trust Sanger Institute, United Kingdom

Western blot - PSD95 antibody - Synaptic Marker (ab18258)

All lanes : Anti-PSD95 antibody - Synaptic Marker (ab18258) at 1 µg/ml

Lane 1 : Brain (Mouse) Tissue Lysate (ab27253)
Lane 2 : Brain (Rat) Tissue Lysate (ab7942)

Lysates/proteins at 10 µg per lane.

Secondary
IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) (ab28446) at 1/10000 dilution

Performed under reducing conditions.

Predicted band size : 80 kDa
Observed band size : 85 kDa (why is the actual band size different from the predicted?)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - PSD95 antibody - Synaptic Marker (ab18258)

ab18258 (1/1000) staining PSD9 in paraffin-embedded rat cerebellum. Tissue was fixed in formaldehyde, blocking performed using 1% BSA (10 mins/RT) and heat mediated antigen retrieval performed before staining. The secondary antibody (1/200) was goat anti rabbit IgG conjugated to Biotin. For further experimental details please refer to abreview.

Carl Hobbs, King`s College London, United Kingdom

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - PSD95 antibody - Synaptic Marker (ab18258)

ab18258 (1/500) staining PSD95 in paraffin-embedded Mouse Cerebellum (Top) and Medulla (Bottom) tissue, showing positive staining to the synaptic regions of the brain.  Tissue was fixed in formaldehyde, blocking performed using 1% BSA (10 mins/RT) and heat mediated antigen retrieval performed before staining. The secondary antibody (1/200) was goat anti rabbit IgG conjugated to Biotin.  For further experimental details please refer to abreview.

This image is courtesy of an abreview submitted by Carl Hobbs

Immunohistochemistry (Frozen sections) - Anti-PSD95 antibody - Synaptic Marker (ab18258)

ab18258 staining PSD95 in murine retinal tissue by Immunohistochemistry (Frozen sections).

Tissue was fixed with paraformaldehyde, blocked using 10% serum for 30 minutes at 24°C, then incubated with ab18258 at a 1/1000 dilution for 16 hours at 4°C. The secondary used was an Alexa Fluor 488 conjugated goat anti-rabbit polyclonal used at a 1/1000 dilution.

Image courtesy of an anonymous Abreview.

Immunohistochemistry (Frozen sections) - Anti-PSD95 antibody - Synaptic Marker (ab18258)

ab18258 staining PSD95 in rat retinal tissue by Immunohistochemistry (Frozen sections).

Tissue was fixed with paraformaldehyde, blocked using 10% serum for 30 minutes at 24°C, then incubated with ab18258 at a 1/1000 dilution for 16 hours at 4°C. The secondary used was an Alexa Fluor 488 conjugated goat anti-rabbit polyclonal used at a 1/1000 dilution.

Image courtesy of an anonymous Abreview.

Immunocytochemistry/ Immunofluorescence - Anti-PSD95 antibody - Synaptic Marker (ab18258)

ab18258 staining PSD95 in human SH-SY5Y cells by Immunocytochemistry/ Immunofluorescence.

Cells were fixed in paraformaldehyde, blocked with 10% serum for 20 minutes at 24°C, then incubated with ab18258 at a 1/1000 dilution for 16 hours at 4°C. The secondary used was an Alexa-Fluor 488 conjugated donkey anti-rabbit polyclonal used at a 1/1000 dilution. Counterstained with Hoechst 33258 (blue).

Image courtesy of an anonymous Abreview.

Immunocytochemistry/ Immunofluorescence - PSD95 antibody - Synaptic Marker (ab18258)

ICC/IF image of ab18258 stained human SHSY5Y cells. The cells were 4% PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab18258, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).