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

1-10 of 13 Q&A

Answer


The antibodies mentioned above are suitable for Rat as well as human samples. ab9018 is yet to be tested in IHC-Fr however as it is tested in IHC-FoFr so it is more likely to work.

The suitable secondary's are for ab4648, ab82259 and ab9018 are

https://www.abcam.com/rabbit-polyclonal-secondary-antibody-to-mouse-igg-hl-hrp-ab97046.html

https://www.abcam.com/goat-polyclonal-secondary-antibody-to-mouse-igg-hl-hrp-ab97023.html

for ab18258 it is

https://www.abcam.com/goat-polyclonal-secondary-antibody-to-rabbit-igg-hl-hrp-ab6721.html

for ab5392 it is;

https://www.abcam.com/goat-polyclonal-secondary-antibody-to-chicken-igy-h-l-hrp-ab97135.html

The protocols are:

https://www.abcam.com/index.html?pageconfig=resource&rid=11417

https://www.abcam.com/index.html?pageconfig=resource&rid=11383

https://www.abcam.com/index.html?pageconfig=resource&rid=13046

IHC-Fr is immunohistochemistry on frozen section

IHC-FoFr is PFA perfusion fixed frozen sections.

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Answer

Thank you for contacting us.

This antibody has been tested positively with different tissue sections but Rat spinal cord. We test our antibodies with as many tissue sections as we can however it is not always possible to test each and every tissue. If the antibody reacts in a particular species then we would assume that it will detect the protein in all the tissue where protein is expressed naturally.

This antibody has been tested in Rat and IHC-Fr so it is fully guaranteed. We will refund your money if it does not work.

Please note we recommend using the positive control with all our antibodies which is a good way to compare, whether the problem is due to antibody or tissue section.

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

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Answer

I am very pleased to hear you would like to test ab5076, ab8049, ab18258, ab58802, ab64581, ab82259 and send us your results. The testing discount doesn't apply to ab4674 since this has already been tested in Rhesus monkey and IHC so we would guarantee that experiment to work.

The codes will give you a value off your next orders before the expiration date. To redeem this offer, please submit 6 Abreview with rhesus monkey data and include the code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.
For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.

The code will be active once the Abreview wit has been submitted and can be redeemed in one of the following ways:
1) Call to place your order and mention the code to our customer service department;
2) Include the code in your fax order;
3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated. Remember if the product does not work you will be covered by our Abpromise guarantee and eligible for a full refund or replacement.

If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: https://www.abcam.com/abtrial.

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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

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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.

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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.

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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

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Question
Answer

I'm happy to help! Please let me know if you have any other questions.

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Answer

Thank you for your enquiry.

After storing this antibod in the freezer for 1 year it should still be fine.

You might try increasing the concentration of the primary two-fold to see if adding more antibody enhances the signal.

I hope this is helpful. Please contact us again if you have any further questions,.

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Answer

Thank you very much for your interest in ab18258.

To our knowledge, ab18258 has not been tested in chicken and we do not have any anti-PSD95 antibodies that have been tested in chicken as far as we know. Therefore, I can offer a discount off a future purchase if you buy ab18258 now, test it in chicken and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of: 1 freeprimary antibody. If you are interested in this offer, please follow these steps:

1. Reply to this e-mail to let me know that you would like to proceed and test ab18258 in chicken. I will then send a discount code. This code must be issued before purchasing ab18258 so please wait for my reply before ordering.

2. Purchase ab18258 either by phone, fax, or online (www.abcam.com).

3. Test it in chicken.

4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any primary antibody ordered and the discount code is valid for 4 months after issue.

We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab18258 turns out to be unsuitable for chicken, you will still receive the discount on your next purchase after your Abreview has been submitted.

Please let me know if you have any questions about this offer and I would be happy to help you further.

The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

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1-10 of 13 Q&A

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