Anti-PSD95 antibody [6G6-1C9] (ab2723)

  Flow Cytometry - PSD95 antibody [6G6-1C9] (ab2723)

Overlay histogram showing SH-SY5Y cells stained with ab2723 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab2723, 1µg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.
Please note that Abcam do not have data for use of this antibody on non-fixed cells. We welcome any customer feedback.

  Western blot - PSD95 antibody [6G6-1C9] (ab2723)

All lanes : Anti-PSD95 antibody [6G6-1C9] (ab2723) at 5 µg/ml

Lane 1 : Hippocampus (Rat) Tissue Lysate
Lane 2 : Forebrain (Rat) Tissue Lysate
Lane 3 : Hippocampus (Mouse) Tissue Lysate
Lane 4 : Forebrain (Mouse) Tissue Lysate

Lysates/proteins at 20 µg per lane.

Secondary
Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed (ab97040) at 1/5000 dilution

Performed under reducing conditions.

Predicted band size : 80 kDa
Observed band size : 95 kDa (why is the actual band size different from the predicted?)
Additional bands at : 100 kDa,110 kDa,80 kDa. We are unsure as to the identity of these extra bands.

Exposure time : 3 minutes

  Immunocytochemistry/ Immunofluorescence - PSD95 antibody [6G6-1C9] (ab2723)

ab2723 at 1/200 staining rat primary hippocampal neurons by ICC/IF. The cells were paraformaldehyde fixed and then permeabilized with 0.1% Triton X-100 before being stained with the antibody for 1 hour. An Alexa-Fluor ® 594 conjugated goat polyclonal antibody was used as the secondary.

The antibody shows numerous puncta along the shafts.

This image is courtesy of an Abreview submitted by Dr Farida Korobova

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  Western blot - PSD95 antibody [6G6-1C9] (ab2723)

Predicted band size : 80 kDa


All lanes : PSD95 antibody (ab2723) at 1/2000 dilution (in blocking buffer for 16 hours at 4°C) + whole tissue lysate of Mouse hippocampus at 10µg

Secondary
An HRP-conjugated Horse anti-mouse IgG polyclonal at 1/10000 dilution
developed using the ECL technique

Performed under reducing conditions

Exposure time : 30 seconds

Blocking Step : 5% Milk for 1 hour at 25°C

This image is courtesy of an anonymous Abreview

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