Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
(Peptide available as ab226770)
WB: SH-SY5Y treated with alkaline phosphatase, Human and Mouse brain tissue lysate;IHC-P: human glioblastoma, human Alzheimer hippocampus, human, mouse and rat colon;IP- Human brain lysate;IHC-Fr: Mouse and Rat cerebrum tissue, Hu Alzheimer brain.
常规说明
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 productionFor 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.
存放说明 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.
存储溶液 pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 59% PBS, 0.05% BSA
The Abpromise guarantee
Abpromise™承诺保证使用ab109390于以下的经测试应用
\"应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。
1/100.
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20)
IHC-P (5) 1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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.
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.
组织特异性 Expressed in neurons. Isoform PNS-tau is expressed in the peripheral nervous system while the others are expressed in the central nervous system.
疾病相关 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.
发展阶段 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.
The tau/MAP repeat binds to tubulin. Type I isoforms contain 3 repeats while type II isoforms contain 4 repeats.
翻译后修饰 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.
细胞定位 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.
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
Immunohistochemistry analysis offrozen mouse cerebrum tissue sections labeling Tau (phospho S396) withab109390 at 1/100 (1 g/mL). ab150077 AlexaFluor 488 Goat anti-Rabbit at1/1000 (2 g/mL) was used as the secondary antibody. Sections were fixed with 4% PFA and permeabilised with 0.2% Triton X-100.DAPI (blue) was used as nuclear counterstain. Antigen retrieval was heat mediated using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
Cytoplasmic staining on mouse cerebrum, the signal decreased after phosphatase treatment at 37℃ for 2h.
Immunohistochemistry analysis of paraffin-embedded human colontissue sections labeling Tau (phospho S396) withab109390 at 1/4000 dilution (0.027 g/mL). Goat Anti-Rabbit IgG H L (HRP polymer) was used as the secondary antibody. Sections were counterstained with Hematoxylin. Antigen retrieval was heat mediated using ab93684 (Tris/EDTA buffer, pH 9.0).
Positive staining on ganglions of human colon without alkaline phosphatase treatment (image A). No staining on ganglions of human colon with alkaline phosphatase treatment (image B).
The section was incubated with ab109390 overnight at +4 C.
Immunohistochemistry analysis of paraffin-embeddedrat colontissue sections labeling Tau (phospho S396) withab109390 at 1/4000 dilution (0.027 g/mL). Goat Anti-Rabbit IgG H L (HRP polymer) was used as the secondary antibody. Sections were counterstained with Hematoxylin. Antigen retrieval was heat mediated using ab93684 (Tris/EDTA buffer, pH 9.0).
Positive staining on ganglions of mouse colon without alkaline phosphatase treatment (image A). No staining on ganglions of mouse colon with alkaline phosphatase treatment (image B).
The section was incubated with ab109390 overnight at +4 C.
Immunohistochemistry analysis of paraffin-embedded rat colontissue sections labeling Tau (phospho S396) with ab109390 at 1/4000 dilution (0.027 g/mL). Goat Anti-Rabbit IgG H L (HRP polymer) was used as the secondary antibody. Sections were counterstained with Hematoxylin. Antigen retrieval was heat mediated using ab93684 (Tris/EDTA buffer, pH 9.0).
Positive staining on ganglions of rat colon without alkaline phosphatase treatment (image A). No staining on ganglions of rat colon with alkaline phosphatase treatment (image B).
The section was incubated with ab109390 overnight at +4 C.
All lanes : Anti-Tau (phospho S396) antibody [EPR2731] (ab109390) at 1/1000 dilution
Lane 1 : Human brain lysates
Lane 2 : Human brain lysates and the membrane was incubated with alkaline phosphatase
Lane 3 : Human brain lysates and the membrane was incubated with lambda phosphatase
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 79 kDa
Observed band size: 50-79 kDa why is the actual band size different from the predicted?
Exposure time: 100 seconds
Blocking/Diluting buffer and concentration 5% NFDM/TBST
Tau assembles into oligomers as described in PMID: 28382304, 32692785 and 30120733.
All lanes : Anti-Tau (phospho S396) antibody [EPR2731] (ab109390) at 1/1000 dilution
Lane 1 : Mouse brain lysates
Lane 2 : Mouse brain lysates and the membrane was incubated with alkaline phosphatase
Lane 3 : Mouse brain lysates and the membrane was incubated with lambda phosphatase
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 79 kDa
Observed band size: 50-79 kDa why is the actual band size different from the predicted?
Exposure time: 10 seconds
Diluting/Diluting buffer and concentration 5% NFDM/TBST
Tau assembles into oligomers as described in PMID: 28382304, 32692785 and 30120733..
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
IHC image of Tau (phospho S396) staining in a section of frozen normalhumanAlzheimer brainperformed 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.
Immunohistochemical staining of paraffin embedded humanglioblastoma 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.
ab109390 at 1/20 immunoprecipitating Tau (phospho S396)inHuman brain lysate.
Lane 1 (input):Human brain lysate(10 g)
Lane 2 (+): ab109390 +Human brain lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab109390inHuman brainlysate.
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 (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 unpurifiedab109390 at 1/1000 dilution for 2 hoursat 21 C. 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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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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ab109390 被引用在 65 文献中.
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 View all Publications for this productBlocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) Concentration: 2% Temperature: 25 C
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) Concentration: 5% Temperature: 24 C
Blocking step Serum as blocking agent for 30 minute(s) Concentration: 5% Temperature: 25 C
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) Concentration: 2% Temperature: 18 C
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) Concentration: 20% Temperature: rt C
Sample Human Cell lysate - whole cell (human stem cell derived neurons (healthy/diseased))
Blocking step BSA as blocking agent for 30 minute(s) Concentration: 5% Temperature: rt C
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