Overview

  • Product name
    Anti-PSD95 antibody [7E3-1B8]
    See all PSD95 primary antibodies
  • Description
    Mouse monoclonal [7E3-1B8] to PSD95
  • Host species
    Mouse
  • Tested applications
    Suitable for: WB, ICC/IF, IHC-P, IHC-Fr, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human
  • Immunogen

    Recombinant full length protein corresponding to Rat PSD95.

  • Positive control
    • ICC/IF: Rat primary hippocampal neurons; HaCaT cells; mouse backskin tissue section. WB: Rat brain membrane lysate; bovine brain tissue extract. Flow Cyt: K562 cells. IHC-Fr: Mouse retinal sections.

Properties

Applications

Our Abpromise guarantee covers the use of ab13552 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
WB Use a concentration of 1 µg/ml. Detects a band of approximately 100 kDa.
ICC/IF Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IHC-Fr Use at an assay dependent concentration. PubMed: 23209820
Flow Cyt Use 1µg for 106 cells.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

 

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

  • Central Panel: ab13552 staining PSD95 in rat primary hippocampal neurons by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.3% Triton and blocked with 3% BSA for 15 minutes at 20°C. Samples were incubated with primary antibody (1/200 in PBS-BSA 3%) for 30 minutes at 20°C. An Cy5®-conjugated Donkey polyclonal to mouse IgG (dilution 1/500) was used as secondary antibody.
    Left-hand panel: Homer GFP.
    Right-hand panel: Merge.

    See Abreview

  • Immunocytochemistry/ Immunofluorescence analysis of HaCaT cells labeling PSD95 with ab13552 at 1/100 dilution. Cells were fixed with 00% cold methanol for 10 minutes at -20°C. A FITC Goat Anti-Mouse antibody at 1/50 was used as secondary. The image shows Filamentouslike staining. 
  • Immunohistochemistry analysis of rat neocortex tissue labeling PSD95 at 1/1000 dilution.

  • Anti-PSD95 antibody [7E3-1B8] (ab13552) at 1/1000 dilution + Rat brain membrane lysate.
  • Western blot analysis of bovine brain tissue extract, probed with ab13552. Western blot analysis of bovine brain tissue extract, probed with ab13552.
  • ab13552 at 1/100 dilution staining PSD95 in mouse backskin tissue section by Immunohistochemistry (Bouin's fixed paraffin embedded tissue sections). Tissue underwent heat mediated antigen retrieval in microwave with 2, 5 minutes incubation intervals in citrate buffer. The image show basal, hair follicle and dermal staining.

  • Overlay histogram showing K562 cells stained with ab13552 (red line). The cells were fixed with 80% methanol (5 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 (ab13552, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
    Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.
  • Immunohistochemical analysis of frozen mouse retinal sections, detecting photoreceptor synaptic endings by staining PSD95 (red) with ab13552.

    Sections were fixed in 4% paraformaldehyde before incubating with primary antibody (1/500). An AlexaFluor®568-conjugated anti-mouse IgG (1/1000) was used as the secondary antibody.

References

This product has been referenced in:
  • Krukowski K  et al. Temporary microglia-depletion after cosmic radiation modifies phagocytic activity and prevents cognitive deficits. Sci Rep 8:7857 (2018). Read more (PubMed: 29777152) »
  • Koshida R  et al. Myosin Id localizes in dendritic spines through the tail homology 1 domain. Exp Cell Res 367:65-72 (2018). WB, ICC . Read more (PubMed: 29559226) »
See all 13 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Application
Immunohistochemistry free floating
Sample
Mouse Tissue sections (Brain)
Specification
Brain

Abcam user community

Verified customer

Submitted Aug 30 2017

Answer

Thank you for your reply.

I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful. Before deciding how to proceed, I would like to investigate this particular case further for you, and also obtain some further information for our quality records.

In order to do this, I have enclosed a questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily. I have completed the sections for which information has already been provided.

In the event that we cannot get the antibody functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund. We are happy to provide an alternative if requested if we are not able to get the antibody working for you.

Thank you for your time and cooperation. We look forward to receiving the completed questionnaire.

Order Details

Antibody code:

Lot number:

Purchase order number
or preferably Abcam order number:


General Information
Antibody storage conditions (temperature/reconstitution etc)


Description of the problem (high background, low signal, non-specific satining etc.)

Sample (Species/Tissue/Cell Type/Cell Line etc.)


Fixation of sample (Ethanol/Methanol/Acetone/Paraformaldehyde/Other/Duration etc.)


Antigen retrieval (Enzymatic method, Heat mediated technique etc.)


Permeabilization step


Blocking conditions (Buffer/time period, Blocking agent etc.)


Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Detection method


Positive and negative controls used (please specify)


Optimization attempts (problem solving)
How many times have you tried the IHC?



Have you run a "No Primary" control?
Yes No

Do you obtain the same results every time?
Yes No


What steps have you altered?


Additional Notes


We would appreciate if you are also able to provide any image which woudl help us to assess the results. From the lot that has worked, and the lot that has not.

Read More

Answer

Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this lot of antibody.

In answer to your question, I would like to reassure you that we monitor feedback closely on a weekly basis and we are not currently concerned about the general quality of this antibody or this batch.

I would also like to reassure you that this antibody is tested and covered by our 6 month guarantee for WB, ICC/IF, IHC-P, Flow Cyt and in use, Rat, Cow and Human samples. In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.

I would like to investigate this particular case further for you, and also obtain more information for our quality monitoring records. In order to proceed with this, I would like to send you a technical questionnaire. This will help you put the information we require together very easily. I would appreciate if you could confirm which application you have been using, so I can send the correct questionnaire.

I would appreciate if you are able to provide any image which would help us to assess the results.

Thank you for your time and cooperation. We look forward to hearing from you with the requested details.

Read More

Answer

Thank you for confirming these details and for your cooperation.

I am sorry ab13552 did not perform well enough for your experiments and for the inconvenience this has caused. But I am happy that we have an alternative available that hopefully works for you. As requested, I have issued a free of charge replacement with ab12093 with the order number x.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

Read More

Answer

Thank you for your answer.

Unfortunately, all the PSD-95 antibodies that we can guarantee for IHC-FoFr are rabbit polyclonals (ab16493, ab16495 and ab18258).

Having said this, we have goat polyclonal anti-PSD-95 antibody ab12093 available which has been tested for IHC on paraffin-embedded sections (IHC-P) and is working for free-floating sections (IHC-FrFl) - details about the IHC-FrFl protocol can be found in the publication by Koffie et al (PMID:19228947, attached). These applications may be closer to your experimental settings and thus this antibody might work for you. If you are happy to try it with this one, I would be pleased to arrange the replacement for you.
https://www.abcam.com/index.html?datasheet=12093 https://www.abcam.com/index.html?datasheet=12093.

Also, we have the mouse monoclonal antibody ab99009 available which is guaranteed to work on paraffin-embedded sections as well as on frozen sections (IHC-P and -Fr), and thus there is a chance that it might work for your experimental layout. If you prefer this one, I could send it to you.
https://www.abcam.com/index.html?datasheet=99009 https://www.abcam.com/index.html?datasheet=99009.

However, I do not want to talk you into using these antibodies, and if none of the above is an option for you, I would be pleased to send either any other primary antibody of your choice or issue a credit note.

I look forward to hearing from you with details of how you would like to proceed.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Primary hippocampal neurons)
Specification
Primary hippocampal neurons
Fixative
Paraformaldehyde
Permeabilization
Yes - Triton 0.3%
Blocking step
BSA as blocking agent for 15 minute(s) · Concentration: 3% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Mar 11 2010

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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