Overview

  • Product name
    Anti-PSD95 antibody [6G6-1C9]
    See all PSD95 primary antibodies
  • Description
    Mouse monoclonal [6G6-1C9] to PSD95
  • Host species
    Mouse
  • Specificity

    We do not guarantee IHC-P for mouse.

  • Tested applications
    Suitable for: IHC-P, WB, IHC-Fr, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Zebrafish
    Predicted to work with: Cow
  • Immunogen

    Recombinant full length protein corresponding to Rat PSD95.

  • Positive control
    • In Western Blot, this antibody gave a positive signal in rat and mouse forebrain and hippocampus tissue lysates. In IHC-P, rat retinal FFPE samples were used as positive control.
  • General notes

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

    The Protocols tab contains a Mouse on Mouse staining protocol with recommendations when using a mouse monoclonal antibody to stain mouse tissues and tips for reducing background.

Properties

Applications

Our Abpromise guarantee covers the use of ab2723 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-P Use a concentration of 1 - 2 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

We do not guarantee IHC-P for mouse.

WB Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 95 kDa (predicted molecular weight: 80 kDa).
IHC-Fr Use at an assay dependent concentration.
IP Use at an assay dependent concentration.

Target

  • Function
    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.
  • Tissue specificity
    Brain.
  • Sequence similarities
    Belongs to the MAGUK family.
    Contains 1 guanylate kinase-like domain.
    Contains 3 PDZ (DHR) domains.
    Contains 1 SH3 domain.
  • 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.
  • Post-translational
    modifications
    Palmitoylation of isoform 1 is required for targeting to postsynaptic density.
  • Cellular localization
    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.
  • Information by UniProt
  • Database links
  • Alternative names
    • Discs large homolog 4 antibody
    • Disks large homolog 4 antibody
    • DLG 4 antibody
    • Dlg4 antibody
    • DLG4_HUMAN antibody
    • FLJ97752 antibody
    • FLJ98574 antibody
    • Human post synaptic density protein 95 antibody
    • Post synaptic density protein 95 antibody
    • Postsynaptic density protein 95 antibody
    • PSD 95 antibody
    • PSD-95 antibody
    • PSD95 antibody
    • SAP 90 antibody
    • SAP-90 antibody
    • SAP90 antibody
    • Synapse associated protein 90 antibody
    • Synapse-associated protein 90 antibody
    • Tax interaction protein 15 antibody
    see all

Images

  • IHC image of ab2723 staining in rat retinal formalin fixed paraffin embedded tissue section, performed on a Leica BONDTM 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 ab2723, 2µg/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. Secondary-only control image is shown as insert.

    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.

  • Lanes 1-6 : Anti-PSD95 antibody [6G6-1C9] (ab2723) at 1/1200 dilution
    Lanes 7-12 : Anti-PSD95 antibody [6G6-1C9] (ab2723) at 1/1200 dilution (18 hours at 4°C)

    Lanes 1-6 : Mouse Hippocampus with Li-Cor Block Buffer, 1 hour at 21°C
    Lanes 7-12 : Mouse Liver with Li-Cor Block Buffer, 1 hour at 21°C

    Lysates/proteins at 30 µg per lane.

    Secondary
    All lanes : Li-Cor IRDye® Donkey anti-mouse 680LT at 1/18000 dilution

    Performed under reducing conditions.

    Predicted band size: 80 kDa
    Additional bands at: 95 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 5 minutes

    See Abreview

  • ab2723 staining cultured rat primary hippocampal neurons by ICC/IF. The cultured neurons were fixed with 4% formaldehyde for 5 minutes and blocked with 10% donkey serum in 0.1% PBS- 0.3% TritonX for 30 minutes at 24°C. The cultured neurons were then stained with ab2723 at 1/1000 in 0.3% TritonX with 0.1x PBS and 10% donkey serum for 4h at 24°C. An Alexa Fluro 568 donkey anti-mouse polyclonal antibody at 1/1000 was used as the secondary antibody. PSD95 (which stains the axons and dendrytic spines) can be observed in red (Alexa fluro 568 secondary). Synaptophysin is observable in green (alexa 488 secondary) and colocalizes with PSD95 in dendritic spines. Nuclei are stained in blue with 1.43µM Hoechst.

    See Abreview

  • 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
    All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (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
  • ab2723 staining PSD95 in Zebrafish retina (outer 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® 647-conjugated goat anti-mouse IgG polyclonal (1/1000) was used as the secondary antibody.

    See Abreview

  • 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.

    See Abreview

  • 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

    See Abreview

  • IHC-FoFr staining of PSD95 on rat hippocampus section using ab2723 (1:3000). The sections used came from animals perfused fixed with Paraformaldehyde 4% with 15% of a solution of saturated picric acid, in phosphate buffer 0.1M. Following postfixation in the same fixative overnight, the brains were cryoprotected in sucrose 30% overnight. Brains were then cut using a cryostat and the immunostainings were performed using the ‘free floating’ technique.

    See Abreview

References

This product has been referenced in:
  • Merino-Serrais P  et al. 27-Hydroxycholesterol Induces Aberrant Morphology and Synaptic Dysfunction in Hippocampal Neurons. Cereb Cortex 29:429-446 (2019). Read more (PubMed: 30395175) »
  • Graham K  et al. Region-specific inhibition of 14-3-3 proteins induces psychomotor behaviors in mice. NPJ Schizophr 5:1 (2019). Read more (PubMed: 30643138) »
See all 90 Publications for this product

Customer reviews and Q&As

1-10 of 17 Abreviews or Q&A

Application
Western blot
Sample
Mouse Tissue lysate - whole (Frontal cortex)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 µg
Specification
Frontal cortex
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 23°C

Dr. Alexandre Magno

Verified customer

Submitted Jun 26 2019

Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse brain primary culture)
Gel Running Conditions
Reduced Denaturing (Reduced Denaturing (NuPAGE 4-12% Bis-Tris precast protein gel))
Loading amount
400000 cells
Specification
Mouse brain primary culture
Blocking step
Li-Cor Odyssey blocking buffer as blocking agent as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 100% · Temperature: 28°C

Abcam user community

Verified customer

Submitted Jan 10 2019

Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (primary hippocampal neurons)
Permeabilization
Yes - 0.2% TritonX 100
Specification
primary hippocampal neurons
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
Fixative
Paraformaldehyde

Dr. Bhavin Shah

Verified customer

Submitted Apr 09 2018

Application
Immunohistochemistry (Frozen sections)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Sample
Zebrafish Tissue sections (retina, outer plexiform layer)
Specification
retina, outer plexiform layer
Permeabilization
Yes - triton X
Fixative
Paraformaldehyde

Dr. Ryan Macdonald

Verified customer

Submitted Oct 02 2014

Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing (8% in TGS)
Sample
Mouse Tissue lysate - whole (Hippocampus and Liver)
Specification
Hippocampus and Liver
Blocking step
Li-Cor Block Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 21°C

Grant Corbett

Verified customer

Submitted Jul 17 2014

Application
Immunoprecipitation
Immuno-precipitation step
Protein A/G
Sample
Mouse Tissue lysate - whole (mouse brain tissue)
Specification
mouse brain tissue
Total protein in input
200 µg

Abcam user community

Verified customer

Submitted Sep 19 2013

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing
Sample
Mouse Tissue lysate - whole (mouse hippocampus and cerebellum)
Specification
mouse hippocampus and cerebellum
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Sep 19 2013

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Rat Tissue sections (Brain)
Specification
Brain
Fixative
Paraformaldehyde
Antigen retrieval step
None
Permeabilization
No

Dr. Sophie Pezet

Verified customer

Submitted Jul 04 2012

Question
Answer

We do not know how PSD95 expression would be affected if toxicity is increased in cells. We encourage customers to consult the latest literature available through PubMed and other resources in order to find the most up-to-date information about their specific research interests.

Read More

Answer

Thank you for contacting Abcam.

In mouse neurons an alternative to the smaller housekeeping proteins such as beta-actin, or GAPDH commonly used as loading control would be to use synaptic markers such as PSD95 or Piccolo.

In western blot I would recommend use of anti-PSD95 antibodieshttps://www.abcam.com/PSD95-antibody-Synaptic-Marker-ab18258.htmlorhttps://www.abcam.com/PSD95-antibody-6G6-1C9-ab2723.htmlwhich will give you band above your threshold size at 95kDA.

Should you want bands with an even higher molecular weight Piccolo has a predicted size at 540kDA, our producthttps://www.abcam.com/Piccolo-antibody-Synaptic-Marker-ab20664.htmlis, like the above PSD95 antibodies, guaranteed to work on mouse samples in Western blot.



I hope that this information is helpful. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.

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1-10 of 17 Abreviews or Q&A

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