Recombinant 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
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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 MouseRatHumanIHC-P HumanIP HumanWB MouseHuman -
Immunogen
This information is proprietary to Abcam and/or its suppliers.
(Peptide available asab226770) -
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.
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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
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
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 59% PBS, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR2731 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab109390 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
Application | Species |
---|---|
IHC-Fr |
Mouse
Rat
Human
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IHC-P |
Human
|
IP |
Human
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WB |
Mouse
Human
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Application | Abreviews | Notes |
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Dot blot |
1/1000.
|
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IHC-Fr | (2) |
1/100.
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20) |
WB | (2) |
1/10000 - 1/50000. Predicted molecular weight: 79 kDa.Can be blocked with Human Tau (phospho S396) peptide (ab226770).
For unpurified, use 1/10000 - 1/50000. |
IP |
1/10 - 1/100.
|
Notes |
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Dot blot
1/1000. |
IHC-Fr
1/100. Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20) |
WB
1/10000 - 1/50000. Predicted molecular weight: 79 kDa.Can be blocked with Human Tau (phospho S396) peptide (ab226770). For unpurified, use 1/10000 - 1/50000. |
IP
1/10 - 1/100. |
Target
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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
modificationsPhosphorylation 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
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Database links
- Entrez Gene: 4137 Human
- Entrez Gene: 17762 Mouse
- Entrez Gene: 29477 Rat
- Omim: 157140 Human
- SwissProt: P10636 Human
- SwissProt: P10637 Mouse
- SwissProt: P19332 Rat
- Unigene: 101174 Human
see all -
Form
There are 9 isoforms produced by alternative splicing. -
Alternative names
- AI413597 antibody
- AW045860 antibody
- DDPAC antibody
see all
Images
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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
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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.
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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.
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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.
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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.
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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.
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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.
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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 minuteDiluting and blocking buffer: 5% NFDM/TBST
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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
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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.
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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
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (54)
ab109390 has been referenced in 54 publications.
- Dillon GM et al. Acute inhibition of the CNS-specific kinase TTBK1 significantly lowers tau phosphorylation at several disease relevant sites. PLoS One 15:e0228771 (2020). PubMed: 32255788
- Gao L et al. Vps35 Deficiency Impairs Cdk5/p35 Degradation and Promotes the Hyperphosphorylation of Tau Protein in Retinal Ganglion Cells. Invest Ophthalmol Vis Sci 61:1 (2020). PubMed: 31995153
- Leong W et al. PP2A subunit PPP2R2C is downregulated in the brains of Alzheimer's transgenic mice. Aging (Albany NY) 12:6880-6890 (2020). PubMed: 32291379
- Ai PH et al. Paroxetine ameliorates prodromal emotional dysfunction and late-onset memory deficit in Alzheimer's disease mice. Transl Neurodegener 9:18 (2020). PubMed: 32398165
- Lin CI et al. 1,25(OH)2D3 Alleviates Aß(25-35)-Induced Tau Hyperphosphorylation, Excessive Reactive Oxygen Species, and Apoptosis Through Interplay with Glial Cell Line-Derived Neurotrophic Factor Signaling in SH-SY5Y Cells. Int J Mol Sci 21:N/A (2020). PubMed: 32545801