Anti-PSD95 antibody - Synaptic Marker

• Supplier: Abcam
• Catalog: ab18258
• Prices: $404.00
• Sizes: 100 µg
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: IgG
• Applications: IHC-F ICC/IF ICC/IF WB IHC-P ICC/IF IHC-F ELISA IP
• Species Reactivities: Mouse, Rat, Human, Zebrafish, Marmoset
• Antigen: Synthetic peptide conjugated to KLH derived from within residues 50 - 150 of Mouse PSD95
• Description: Rabbit Polyclonal
• Gene: DLG4
• Conjugate: Unconjugated

Product images

IHC image of PSD95 staining in mouse brain formalin fixed paraffin embedded tissue section, performed on a Leica Bond system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab18258, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

All lanes : Anti-PSD95 antibody - Synaptic Marker (ab18258) at 1/350 dilutionLane 1 : Marmoset (common) brain whole tissue lysateLane 2 : Marmoset (common) brain whole tissue lysateLane 3 : Marmoset (common) brain whole tissue lysateLane 4 : Marmoset (common) brain whole tissue lysateLysates/proteins at 25 µg per lane.SecondaryHRP-conjugated goat anti-rabbit IgG polyclonal at 1/12000 dilutionPerformed under reducing conditions.

All lanes : Anti-PSD95 antibody - Synaptic Marker (ab18258) at 1 µg/mlLane 1 : wildtype mouse brain tissue lysateLane 2 : PSD95 knockout mouse brain tissue lysateLysates/proteins at 20 µg per lane.Performed under reducing conditions.

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.

All lanes : Anti-PSD95 antibody - Synaptic Marker (ab18258) at 1 µg/mlLane 1 : Brain (Mouse) Tissue Lysate (ab27253)Lane 2 : Brain (Rat) Tissue Lysate (ab7942)Lysates/proteins at 10 µg per lane.SecondaryRabbit IgG secondary antibody (ab28446) at 1/10000 dilutionPerformed under reducing conditions.

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.See Abreview

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.See Abreview

ab18258 staining PSD95 in Zebrafish retina (inner plexiform layer) tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde, permeabilized with Triton X-100 and blocked with 5% BSA for 1 hour at 23°C. Sodium citrate antigen retrieval was used. Samples were incubated with primary antibody (1/100) for 16 hours at 4°C. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG polyclonal (1/1000) was used as the secondary antibody.See Abreview

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.See Abreview

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.See Abreview

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).See Abreview

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).

Product References

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