Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Tau (phospho S396) antibody [EPR2731] (ab109390)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR2731] to Tau (phospho S396)
  • Suitable for: Dot blot, IHC-Fr, WB, IP
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-Tau (phospho S396) antibody [EPR2731]
    See all Tau primary antibodies

  • Description

    Rabbit monoclonal [EPR2731] to Tau (phospho S396)

  • Host species

    Rabbit

  • Specificity

    This antibody only detects Tau phosphorylated at serine 396.

  • Tested Applications & Species

    Application Species
    IHC-Fr
    Mouse
    Rat
    Human
    IHC-P
    Human
    IP
    Human
    WB
    Mouse
    Human
    See all applications and species data

  • Immunogen

    This information is proprietary to Abcam and/or its suppliers.
    (Peptide available as ab226770)

  • Positive control

    • WB: SH-SY5Y treated with alkaline phosphatase, Mouse brain; ICC/IF: Neuro-2a; IHC-P: human glioblastoma, human Alzheimer hippocampus; IP- Human brain lysate; IHC-Fr: Mouse and Rat cerebrum tissue, Hu Alzheimer brain.

  • General notes

    Tau is a protein associated with several disease states, known collectively as tauopathies. The most well-known of these is Alzheimer’s disease (AD), were tau exhibiting excessive phosphorylation, aggregating to form neurofibrillary tangles. The epitope defined by phosphorylation of S396 in tau is strongly implicated in AD-associated tau pathology, providing a valuable target for the development of therapeutic antibodies to capture tau and prevent spreading of tau pathology. 

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

Images

  • Western blot - Anti-Tau (phospho S396) antibody [EPR2731] (ab109390)
    Western blot - Anti-Tau (phospho S396) antibody [EPR2731] (ab109390)
    All lanes : Anti-Tau (phospho S396) antibody [EPR2731] (ab109390) at 1/20000 dilution (purified)

    Lane 1 : Untreated SH-SY5Y
    Lane 2 : SH-SY5Y treated with alkaline phosphatase

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP goat ant-rabbit (H+L) at 1/1000 dilution

    Predicted band size: 79 kDa
    Observed band size: 50-70 kDa
    why is the actual band size different from the predicted?



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Immunohistochemistry (Frozen sections) - Anti-Tau (phospho S396) antibody [EPR2731] (ab109390)
    Immunohistochemistry (Frozen sections) - Anti-Tau (phospho S396) antibody [EPR2731] (ab109390)

    IHC image of Tau (phospho S396) staining in a section of frozen normal human Alzheimer brain performed on a Leica BONDTM system using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab109390, 1/1000 dilution, 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. The inset secondary-only control image is taken from an identical assay without primary antibody.

    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.

  • Immunohistochemistry (Frozen sections) - Anti-Tau (phospho S396) antibody [EPR2731] (ab109390)
    Immunohistochemistry (Frozen sections) - Anti-Tau (phospho S396) antibody [EPR2731] (ab109390)

    Immunohistochemistry (Frozen sections) analysis of rat cerebrum tissue sections labeling Tau with Purified ab109390 at 1/100 (1.0 µg/ml).Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. DAPI was used as a counterstain.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tau (phospho S396) antibody [EPR2731] (ab109390)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tau (phospho S396) antibody [EPR2731] (ab109390)

    Immunohistochemical staining of paraffin embedded human glioblastoma with purified ab109390 at a dilution of 1/4000. A pre-diluted HRP polymer for rabbit/mouse IgG was used as the secondary antibody and the sample was counter-stained wirh hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

  • Immunoprecipitation - Anti-Tau (phospho S396) antibody [EPR2731] (ab109390)
    Immunoprecipitation - Anti-Tau (phospho S396) antibody [EPR2731] (ab109390)

    ab109390 at 1/20 immunoprecipitating Tau (phospho S396) in Human brain lysate.

    Lane 1 (input): Human brain lysate (10µg)

    Lane 2 (+): ab109390 + Human brain lysate.

    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab109390 in Human brain lysate.

    For western blotting, ab109390  at 1/1000 dilution followed by VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

    Diluting / Blocking buffer and concentration: 5% NFDM/TBST.

  • Immunohistochemistry (Frozen sections) - Anti-Tau (phospho S396) antibody [EPR2731] (ab109390)
    Immunohistochemistry (Frozen sections) - Anti-Tau (phospho S396) antibody [EPR2731] (ab109390)

    Immunohistochemistry (Frozen sections) analysis of mouse cerebrum tissue sections labeling Tau with Purified ab109390 at 1/100 (1.0 µg/ml).Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. DAPI was used as a counterstain.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tau (phospho S396) antibody [EPR2731] (ab109390)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tau (phospho S396) antibody [EPR2731] (ab109390) Image courtesy of Carl Hobbs, Kings College London, U.K.

    IHC image of Tau (phospho S396) staining in human Alzheimer hippocampus formalin fixed paraffin embedded tissue section. The section was pre-treated using heat mediated antigen retrieval with citrate buffer. The section was then incubated with unpurified ab109390 at 1/1000 dilution for 2 hoursat 21ºC. A biotin conjugated goat-anti-rabbit antibody was used as a secondary at 1/250. The section shows clear neurofibrillary tangles in a subset of neurons.

    See Abreview

  • Western blot - Anti-Tau (phospho S396) antibody [EPR2731] (ab109390)
    Western blot - Anti-Tau (phospho S396) antibody [EPR2731] (ab109390)
    All lanes : Anti-Tau (phospho S396) antibody [EPR2731] (ab109390) at 1/20000 dilution

    Lane 1 : Human brain whole cell lysates.
    Lane 2 : Human brain whole cell lysates. The membrane was incubated with phosphatase.

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 79 kDa


    Exposure time: 1 minute


    Diluting and blocking buffer: 5% NFDM/TBST

  • Western blot - Anti-Tau (phospho S396) antibody [EPR2731] (ab109390)
    Western blot - Anti-Tau (phospho S396) antibody [EPR2731] (ab109390)
    Anti-Tau (phospho S396) antibody [EPR2731] (ab109390) at 1/5000 dilution (purified) + Mouse brain at 10 µg

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size: 79 kDa
    Observed band size: 50-70 kDa why is the actual band size different from the predicted?



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Dot Blot - Anti-Tau (phospho S396) antibody [EPR2731] (ab109390)
    Dot Blot - Anti-Tau (phospho S396) antibody [EPR2731] (ab109390)

    Dot blot analysis of Tau (phospho S396) phospho peptide (Lane 1) and Tau non-phospho peptide (Lane 2) labeling Tau (phospho S396) with ab109390 at a dilution of 1/1000. ab97051 (Peroxidase conjugated goat anti-rabbit IgG) (H+L) at 1/100 000 was used as the secondary antibody.

    Blocking and diluting buffer: 5% NFDM/TBST.

    Exposure time: 3 minutes.

  • Western blot - Anti-Tau (phospho S396) antibody [EPR2731] (ab109390)
    Western blot - Anti-Tau (phospho S396) antibody [EPR2731] (ab109390)
    All lanes : Anti-Tau (phospho S396) antibody [EPR2731] (ab109390) at 1/10000 dilution (unpurified)

    Lane 1 : SH-SY5Y cell lysates, untreated
    Lane 2 : SH-SY5Y cell lysates, treated with Alkaline Phosphatase

    Lysates/proteins at 10 µg per lane.

    Predicted band size: 79 kDa

  • Anti-Tau (phospho S396) antibody [EPR2731] (ab109390)
    Anti-Tau (phospho S396) antibody [EPR2731] (ab109390)

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