Recombinant
RabMAb

Recombinant Anti-Tau (phospho S404) antibody [EPR2605] (ab92676)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR2605] to Tau (phospho S404)
  • Suitable for: IP, ICC/IF, WB, IHC-P, IHC-Fr, Dot blot
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-Tau (phospho S404) antibody [EPR2605]
    See all Tau primary antibodies

  • Description

    Rabbit monoclonal [EPR2605] to Tau (phospho S404)

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IP, ICC/IF, WB, IHC-P, IHC-Fr, Dot blotmore details
    Unsuitable for: Flow Cyt

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human Tau aa 350-450 (phospho S404). The exact sequence is proprietary.

  • Positive control

    • WB: SH-SY5Y cell lysate, Human brain tissue lysate; ICC/IF: Mouse primary cortical cultures; IP: Human brain tissue lysate; IHC-Fr: Mouse cerebrum tissue, Hu Alzheimer brain.

  • General notes

    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.

Properties

Applications

Our Abpromise guarantee covers the use of ab92676 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
IP 1/20.
ICC/IF Use at an assay dependent concentration.
WB 1/500 - 1/2000. Predicted molecular weight: 79 kDa.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IHC-Fr 1/50.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20)
Dot blot Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for Flow Cyt.

    • Target

    • Function

      Promotes microtubule assembly and stability, and might be involved in the establishment and maintenance of neuronal polarity. The C-terminus binds axonal microtubules while the N-terminus binds neural plasma membrane components, suggesting that tau functions as a linker protein between both. Axonal polarity is predetermined by tau localization (in the neuronal cell) in the domain of the cell body defined by the centrosome. The short isoforms allow plasticity of the cytoskeleton whereas the longer isoforms may preferentially play a role in its stabilization.

    • Tissue specificity

      Expressed in neurons. Isoform PNS-tau is expressed in the peripheral nervous system while the others are expressed in the central nervous system.

    • Involvement in disease

      Note=In Alzheimer disease, the neuronal cytoskeleton in the brain is progressively disrupted and replaced by tangles of paired helical filaments (PHF) and straight filaments, mainly composed of hyperphosphorylated forms of TAU (PHF-TAU or AD P-TAU).
      Defects in MAPT are a cause of frontotemporal dementia (FTD) [MIM:600274]; also called frontotemporal dementia (FTD), pallido-ponto-nigral degeneration (PPND) or historically termed Pick complex. This form of frontotemporal dementia is characterized by presenile dementia with behavioral changes, deterioration of cognitive capacities and loss of memory. In some cases, parkinsonian symptoms are prominent. Neuropathological changes include frontotemporal atrophy often associated with atrophy of the basal ganglia, substantia nigra, amygdala. In most cases, protein tau deposits are found in glial cells and/or neurons.
      Defects in MAPT are a cause of Pick disease of the brain (PIDB) [MIM:172700]. It is a rare form of dementia pathologically defined by severe atrophy, neuronal loss and gliosis. It is characterized by the occurrence of tau-positive inclusions, swollen neurons (Pick cells) and argentophilic neuronal inclusions known as Pick bodies that disproportionally affect the frontal and temporal cortical regions. Clinical features include aphasia, apraxia, confusion, anomia, memory loss and personality deterioration.
      Note=Defects in MAPT are a cause of corticobasal degeneration (CBD). It is marked by extrapyramidal signs and apraxia and can be associated with memory loss. Neuropathologic features may overlap Alzheimer disease, progressive supranuclear palsy, and Parkinson disease.
      Defects in MAPT are a cause of progressive supranuclear palsy type 1 (PSNP1) [MIM:601104, 260540]; also abbreviated as PSP and also known as Steele-Richardson-Olszewski syndrome. PSNP1 is characterized by akinetic-rigid syndrome, supranuclear gaze palsy, pyramidal tract dysfunction, pseudobulbar signs and cognitive capacities deterioration. Neurofibrillary tangles and gliosis but no amyloid plaques are found in diseased brains. Most cases appear to be sporadic, with a significant association with a common haplotype including the MAPT gene and the flanking regions. Familial cases show an autosomal dominant pattern of transmission with incomplete penetrance; genetic analysis of a few cases showed the occurrence of tau mutations, including a deletion of Asn-613.

    • Sequence similarities

      Contains 4 Tau/MAP repeats.

    • Developmental stage

      Four-repeat (type II) tau is expressed in an adult-specific manner and is not found in fetal brain, whereas three-repeat (type I) tau is found in both adult and fetal brain.

    • Domain

      The tau/MAP repeat binds to tubulin. Type I isoforms contain 3 repeats while type II isoforms contain 4 repeats.

    • Post-translational
      modifications

      Phosphorylation at serine and threonine residues in S-P or T-P motifs by proline-directed protein kinases (PDPK: CDK1, CDK5, GSK-3, MAPK) (only 2-3 sites per protein in interphase, seven-fold increase in mitosis, and in PHF-tau), and at serine residues in K-X-G-S motifs by MAP/microtubule affinity-regulating kinase (MARK) in Alzheimer diseased brains. Phosphorylation decreases with age. Phosphorylation within tau's repeat domain or in flanking regions seems to reduce tau's interaction with, respectively, microtubules or plasma membrane components. Phosphorylation on Ser-610, Ser-622, Ser-641 and Ser-673 in several isoforms during mitosis.
      Polyubiquitinated. Requires functional TRAF6 and may provoke SQSTM1-dependent degradation by the proteasome (By similarity). PHF-tau can be modified by three different forms of polyubiquitination. 'Lys-48'-linked polyubiquitination is the major form, 'Lys-6'-linked and 'Lys-11'-linked polyubiquitination also occur.
      Glycation of PHF-tau, but not normal brain tau. Glycation is a non-enzymatic post-translational modification that involves a covalent linkage between a sugar and an amino group of a protein molecule forming ketoamine. Subsequent oxidation, fragmentation and/or cross-linking of ketoamine leads to the production of advanced glycation endproducts (AGES). Glycation may play a role in stabilizing PHF aggregation leading to tangle formation in AD.

    • Cellular localization

      Cytoplasm > cytosol. Cell membrane. Cytoplasm > cytoskeleton. Cell projection > axon. Mostly found in the axons of neurons, in the cytosol and in association with plasma membrane components.
    • Information by UniProt

    • Database links

      see all

    • Form

      There are 9 isoforms produced by alternative splicing.

    • Alternative names

      • AI413597 antibody
      • AW045860 antibody
      • DDPAC antibody
      • FLJ31424 antibody
      • FTDP 17 antibody
      • G protein beta1/gamma2 subunit interacting factor 1 antibody
      • MAPT antibody
      • MAPTL antibody
      • MGC134287 antibody
      • MGC138549 antibody
      • MGC156663 antibody
      • Microtubule associated protein tau antibody
      • Microtubule associated protein tau isoform 4 antibody
      • Microtubule-associated protein tau antibody
      • MSTD antibody
      • Mtapt antibody
      • MTBT1 antibody
      • MTBT2 antibody
      • Neurofibrillary tangle protein antibody
      • Paired helical filament tau antibody
      • Paired helical filament-tau antibody
      • PHF tau antibody
      • PHF-tau antibody
      • PPND antibody
      • PPP1R103 antibody
      • Protein phosphatase 1, regulatory subunit 103 antibody
      • pTau antibody
      • RNPTAU antibody
      • TAU antibody
      • TAU_HUMAN antibody
      • Tauopathy and respiratory failure antibody
      • Tauopathy and respiratory failure, included antibody
      see all

    Images

    • Western blot - Anti-Tau (phospho S404) antibody [EPR2605] (ab92676)
      Western blot - Anti-Tau (phospho S404) antibody [EPR2605] (ab92676)
      All lanes : Anti-Tau (phospho S404) antibody [EPR2605] (ab92676) at 1/500 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
      Observed band size: 50-70 kDa
      why is the actual band size different from the predicted?


      Exposure time: 15 seconds


      Blocking/Diluting buffer and concentration 5% NFDM/TBST

    • Immunocytochemistry/ Immunofluorescence - Anti-Tau (phospho S404) antibody [EPR2605] (ab92676)
      Immunocytochemistry/ Immunofluorescence - Anti-Tau (phospho S404) antibody [EPR2605] (ab92676)

      Immunocytochemistry/ Immunofluorescence analysis of Embryonic mouse primary neural cells labeling Tau with purified ab92676 at 1:100 dilution (1 μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Goat anti rabbit IgG (Alexa Fluor© 488, ab150077) was used as the secondary antibody at 1:1000 (2 μg/ml) dilution. DAPI (blue) was used as nuclear counterstain. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1:200 (2.5 μg/ml) dilution. DAPI (blue) was used as nuclear counterstain. 

      Confocal image showing positive staining in neuron.

    • Immunohistochemistry (Frozen sections) - Anti-Tau (phospho S404) antibody [EPR2605] (ab92676)
      Immunohistochemistry (Frozen sections) - Anti-Tau (phospho S404) antibody [EPR2605] (ab92676)

      IHC image of Tau (phospho S404) 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 ab92676, 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 S404) antibody [EPR2605] (ab92676)
      Immunohistochemistry (Frozen sections) - Anti-Tau (phospho S404) antibody [EPR2605] (ab92676)

      Immunohistochemistry (Frozen sections) analysis of mouse cerebrum tissue sections labeling Tau with Purified ab92676 at 1/50 (1.9 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 S404) antibody [EPR2605] (ab92676)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tau (phospho S404) antibody [EPR2605] (ab92676)

      ab92676 staining Tau (phospho S404) in Human brain tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Samples were incubated with primary antibody (1/100).

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Immunocytochemistry/ Immunofluorescence - Anti-Tau (phospho S404) antibody [EPR2605] (ab92676)
      Immunocytochemistry/ Immunofluorescence - Anti-Tau (phospho S404) antibody [EPR2605] (ab92676)This image is courtesy of an anonymous Abreview.

      Mouse primary cortical cultures stained for Tau (phospho S404) (green) using ab92676 at 1/100 dilution in ICC/IF, followed by Donkey anti-Rabbit IgG (H+L) Alexa Fluor 488®.

      See Abreview

    • Immunoprecipitation - Anti-Tau (phospho S404) antibody [EPR2605] (ab92676)
      Immunoprecipitation - Anti-Tau (phospho S404) antibody [EPR2605] (ab92676)

      ab92676 at 1/20 immunoprecipitating Tau (phospho S404) in human brain tissue lysate.

      The faint bands detected between 14kDa and 25kDa are cleaved fragments of tau (PMID: 16246844, PMID:9736630).

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



      Lanes 1-2 : Anti-Tau (phospho S404) antibody [EPR2605] (ab92676) at 1/1000 dilution
      Lane 3 : Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

      All lanes : Human brain lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/5000 dilution

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


      Exposure time: 30 seconds
    • Western blot - Anti-Tau (phospho S404) antibody [EPR2605] (ab92676)
      Western blot - Anti-Tau (phospho S404) antibody [EPR2605] (ab92676)
      All lanes : Anti-Tau (phospho S404) antibody [EPR2605] (ab92676) at 1/1000 dilution

      Lane 1 : SH-SY5Y cell lysate (Untreated)
      Lane 2 : SH-SY5Y cell lysate (Treated with Okadaic Acid and Calyculin A)

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size: 79 kDa



      Bottom panel: Beta tubulin loading control.
    • Dot Blot - Anti-Tau (phospho S404) antibody [EPR2605] (ab92676)
      Dot Blot - Anti-Tau (phospho S404) antibody [EPR2605] (ab92676)

      Dot blot analysis of Tau (pS404) phosopho peptide (Lane 1) and Tau non-phospho peptide (Lane 2) labelling Tau (phospho S404) with ab92676 at a dilution of 1/1000. ab97051 (Peroxidase conjugated goat anti-rabbit IgG (H+L)) was used as the secondary antibody at a dilution of 1/100000.

      Blocking and dilution buffer: 5% NFDM/TBST.

      Exposure time: 3 minutes.

    References

    Publishing research using ab92676? Please let us know so that we can cite the reference in this datasheet.

    ab92676 has been referenced in 9 publications.

    • Huang R  et al. Increased Ratio of Global O-GlcNAcylation to Tau Phosphorylation at Thr212 Site Is Associated With Better Memory Function in Patients With Type 2 Diabetes. Front Physiol 10:110 (2019). PubMed: 30837891
    • Di Domenico F  et al. Restoration of aberrant mTOR signaling by intranasal rapamycin reduces oxidative damage: Focus on HNE-modified proteins in a mouse model of down syndrome. Redox Biol N/A:101162 (2019). PubMed: 30876754
    • Paonessa F  et al. Microtubules Deform the Nuclear Membrane and Disrupt Nucleocytoplasmic Transport in Tau-Mediated Frontotemporal Dementia. Cell Rep 26:582-593.e5 (2019). PubMed: 30650353
    • Xiang XJ  et al. Mitochondrial methionine sulfoxide reductase B2 links oxidative stress to Alzheimer's disease-like pathology. Exp Neurol 318:145-156 (2019). PubMed: 31078523
    • Stevens CH  et al. Increased Tau Phosphorylation in Motor Neurons From Clinically Pure Sporadic Amyotrophic Lateral Sclerosis Patients. J Neuropathol Exp Neurol 78:605-614 (2019). PubMed: 31131395
    • Zhu H  et al. GSK3ß-mediated tau hyperphosphorylation triggers diabetic retinal neurodegeneration by disrupting synaptic and mitochondrial functions. Mol Neurodegener 13:62 (2018). PubMed: 30466464
    • Jeong JH & Kang EB Effects of treadmill exercise on PI3K/AKT/GSK-3ß pathway and tau protein in high-fat diet-fed rats. J Exerc Nutrition Biochem 22:9-14 (2018). PubMed: 29673239
    • Stoffel W  et al. SMPD3 deficiency perturbs neuronal proteostasis and causes progressive cognitive impairment. Cell Death Dis 9:507 (2018). WB ; Mouse . PubMed: 29725009
    • Cao L  et al. Pseudo-phosphorylation at AT8 epitopes regulates the tau truncation at aspartate 421. Exp Cell Res 370:103-115 (2018). PubMed: 29908160

    Customer reviews and Q&As

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    1-3 of 3 Abreviews or Q&A

    Western blot abreview for Anti-Tau (phospho S404) antibody [EPR2605]

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Basal Forebrain Cholinergic Neurons)
    Gel Running Conditions
    Reduced Denaturing (8% Acrylamide)
    Loading amount
    5 µg
    Specification
    Basal Forebrain Cholinergic Neurons
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Read More

    Dr. Martin Engel

    Verified customer

    Submitted May 23 2016

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Tau (phospho S404) antibody [EPR2605]

     Good
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (primary cortical cultures)
    Permeabilization
    Yes - 0.25% Triton X-100 in PBS
    Specification
    primary cortical cultures
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 22°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Jan 13 2016

    Western blot abreview for Anti-Tau (phospho S404) antibody [EPR2605]

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    10 µg
    Gel Running Conditions
    Reduced Denaturing (10%)
    Sample
    Human Cell lysate - other (human stem cell derived neurons (healthy/diseased))
    Specification
    human stem cell derived neurons (healthy/diseased)
    Treatment
    quinolinic acid 72h (and non treated)
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: rt°C
    Read More

    Abcam user community

    Verified customer

    Submitted Mar 26 2014

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