Overview

  • Product name

  • Description

    Rabbit polyclonal to PSD95
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-FoFr, ICC/IF, WB, IHC-P, ICC, IHC-Fr, Sandwich ELISA, IPmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human, Zebrafish, Cynomolgus monkey, Common marmoset
  • Immunogen

    Synthetic peptide corresponding to Mouse PSD95 aa 50-150 (internal sequence) conjugated to keyhole limpet haemocyanin (Cysteine residue).
    (Peptide available as ab18661)

  • Positive control

    • WB: Mouse and rat brain lysates. ICC/IF: Zebrafish retina (inner plexiform layer) tissue, SHSY5Y cell line
  • General notes

      

Properties

Applications

Our Abpromise guarantee covers the use of ab18258 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-FoFr Use at an assay dependent concentration. PubMed: 20874778
ICC/IF Use a concentration of 1 µg/ml.
WB Use a concentration of 1 - 3 µg/ml. Detects a band of approximately 85 kDa (predicted molecular weight: 80 kDa).

Abcam recommends using 3% milk as the blocking agent.

IHC-P Use at an assay dependent concentration.
ICC Use at an assay dependent concentration.
IHC-Fr 1/1000. PubMed: 17959791
Sandwich ELISA Use at an assay dependent concentration. PubMed: 20169337Used as detection antibody.
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

  • Immunohistochemical analysis in the CA1 region of hippocampus

    Expression of PSD-95 was investigated in TCblR/CD320 KO mice. PSD-95 expression remained the same in both the KO (n = 2) and in the WT (n = 2) mouse.

    The lower panel shows a magnified image of the indicated region.

    PSD-95 was detected with ab18258 at 1/100 dilution.

    (After Figure 7 of Arora et al).

  • All lanes : Anti-PSD95 antibody (ab18258) at 1/350 dilution

    All lanes : Marmoset (common) brain whole tissue lysate

    Lysates/proteins at 25 µg per lane.

    Secondary
    All lanes : HRP-conjugated goat anti-rabbit IgG polyclonal at 1/12000 dilution

    Performed under reducing conditions.

    Predicted band size: 80 kDa
    Observed band size: 80 kDa


    Exposure time: 16 seconds

    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

  • ab18258 (1/1000) staining PSD9 in paraffin-embedded rat cerebellum. Tissue was fixed in formaldehyde, blocking performed using 1% BSA (10 minutes/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

  • Lane 1 - Input lane 500 µg mouse brain lysate

    Lane 2 - IP lane 50 µg mouse brain lysate

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

  • 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

  • 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 minutes. The section was then incubated with ab18258, 1 µg/ml, for 15 minutes 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 (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
    All lanes : Rabbit IgG secondary antibody (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?

  • All lanes : Anti-PSD95 antibody (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?

  • ab18258 staining PSD95 in SHSY5Y cells. The cells were fixed with 100% methanol (5 min) at room temperature, permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with the antibody ab18258 at 5µg/ml and ab7291 (Mouse monoclonal to alpha Tubulin - Loading Control) used at a 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed, (pseudo-colored green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594) preadsorbed, (colored red), both used at a 1/1000 dilution for 1 hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 µM for 1hour at room temperature.

  • 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 minutes/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 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

References

This product has been referenced in:

  • Yuan C  et al. OAB-14, a bexarotene derivative, improves Alzheimer's disease-related pathologies and cognitive impairments by increasing ß-amyloid clearance in APP/PS1 mice. Biochim Biophys Acta Mol Basis Dis 1865:161-180 (2019). Read more (PubMed: 30389579) »
  • Zhong Q  et al. FCPR16, a novel phosphodiesterase 4 inhibitor, produces an antidepressant-like effect in mice exposed to chronic unpredictable mild stress. Prog Neuropsychopharmacol Biol Psychiatry 90:62-75 (2019). Read more (PubMed: 30391306) »
See all 134 Publications for this product

Customer reviews and Q&As

21-30 of 50 Abreviews or Q&A

Application
Immunocytochemistry
Sample
Human Cell (Kelly Neuroblastoma cell line)
Specification
Kelly Neuroblastoma cell line
Fixative
Paraformaldehyde
Permeabilization
No
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 10% · Temperature: 24°C

Abcam user community

Verified customer

Submitted Sep 28 2012

Answer

Thank you for contacting us. I want to apologize for the delay as we arranged to add each peptide to our catalog.

The blocking peptides which you have requested are now available and can be located at the following links:



ab18258 peptide is: ab18661 https://www.abcam.com/psd95-peptide-ab18661.html

ab12093 peptide is: ab12419 https://www.abcam.com/psd95-peptide-ab12419.html

ab18251: peptide is ab18648 https://www.abcam.com/alpha-tubulin-peptide-ab18648.html





I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Neuro2a)
Loading amount
6 µg
Specification
Neuro2a
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (10%)
Blocking step
Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Aug 22 2012

Application
Western blot
Sample
Rat Cell lysate - whole cell (B50 cell line)
Loading amount
6 µg
Specification
B50 cell line
Gel Running Conditions
Reduced Denaturing (10%)
Blocking step
Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Jul 09 2012

Application
Immunocytochemistry
Sample
Rat Cultured Cells (33B cell line)
Specification
33B cell line
Fixative
Paraformaldehyde
Permeabilization
No
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 24°C

Abcam user community

Verified customer

Submitted Jul 06 2012

Application
Immunocytochemistry
Sample
Mouse Cultured Cells (Neuro2a cell line)
Specification
Neuro2a cell line
Fixative
Paraformaldehyde
Permeabilization
No
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 24°C

Abcam user community

Verified customer

Submitted Jul 06 2012

Answer

Thank you very much for contacting us with your question.

This code did expire on March 23, 2012.However, I've justextended the expiration date for this code, and it needs to be used before July 12, 2012 (one week from today) so please have the order placed before then.

I hope this will be helpful, but please let me know if you have further questions or if there is anything else that we can do for you.

Read More

Answer

Thank you for contacting us.


I am not sure how PSD95 expression would be affected if toxicity is increased in your cells would not wantto recommend a product that may be unsuitable. 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.


I am sorry that I could not be more helpful, but I hope that the available literature in this area can provide some clarification. Please do not hesitate to contact us again with other needs or with any questions about our products.

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.

Use our products? Submit an Abreview. Earn rewards!

https://www.abcam.com/abreviews

Read More
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Neuro2a)
Specification
Neuro2a
Fixative
Paraformaldehyde
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 10% · Temperature: 24°C

Abcam user community

Verified customer

Submitted Jun 07 2012

21-30 of 50 Abreviews or Q&A

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