Anti-Tau (phospho S396) 抗体 [E178] (ab32057)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [E178] to Tau (phospho S396)
- Suitable for: IHC-P, Dot blot, ELISA, IHC-Fr, WB, IP
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
製品の概要
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製品名
Anti-Tau (phospho S396) antibody [E178]
Tau 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [E178] to Tau (phospho S396) -
由来種
Rabbit -
特異性
The specificity of this antibody refers to P10636-8.
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アプリケーション
適用あり: IHC-P, Dot blot, ELISA, IHC-Fr, WB, IPmore details
適用なし: Flow Cyt -
種交差性
交差種: Mouse, Rat, Human
交差が予測される動物種: Cow -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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ポジティブ・コントロール
- WB: Human and mouse brain tissue lysates; IP: Human fetal brain lysates; IHC-P: Human cerebrum and salivary gland; Mouse colon and rat colon and tongue tissue; IHC-Fr: Mouse and Rat cerebrum tissue, Human Alzheimer brain tissue
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特記事項
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.
製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
解離定数(KD 値)
KD = 2.08 x 10 -11 M Learn more about KD -
バッファー
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
E178 -
アイソタイプ
IgG -
研究分野
関連製品
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Alternative Versions
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Compatible Secondaries
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Isotype control
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab32057の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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IHC-P | (1) |
1/1000 - 1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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AP |
Use at an assay dependent concentration.
Antibody concentration range - 3.33, 1.67, 0.83, 0.42, 0.21, 0 nM/mL |
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Dot blot |
1/1000.
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ELISA |
Use at an assay dependent concentration.
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IHC-Fr | (1) |
1/500.
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20) |
WB | (2) |
1/1000. Predicted molecular weight: 79 kDa.
For unpurified use at 1/5000 |
IP |
1/20.
For unpurified use at 1/100 |
特記事項 |
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IHC-P
1/1000 - 1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
AP
Use at an assay dependent concentration. Antibody concentration range - 3.33, 1.67, 0.83, 0.42, 0.21, 0 nM/mL |
Dot blot
1/1000. |
ELISA
Use at an assay dependent concentration. |
IHC-Fr
1/500. Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20) |
WB
1/1000. Predicted molecular weight: 79 kDa. For unpurified use at 1/5000 |
IP
1/20. For unpurified use at 1/100 |
ターゲット情報
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機能
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. -
組織特異性
Expressed in neurons. Isoform PNS-tau is expressed in the peripheral nervous system while the others are expressed in the central nervous system. -
関連疾患
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. -
配列類似性
Contains 4 Tau/MAP repeats. -
発生段階
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. -
ドメイン
The tau/MAP repeat binds to tubulin. Type I isoforms contain 3 repeats while type II isoforms contain 4 repeats. -
翻訳後修飾
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. -
細胞内局在
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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参照データベース
- Entrez Gene: 281296 Cow
- Entrez Gene: 4137 Human
- Entrez Gene: 17762 Mouse
- Entrez Gene: 29477 Rat
- Omim: 157140 Human
- SwissProt: P29172 Cow
- SwissProt: P10636 Human
- SwissProt: P10637 Mouse
see all -
製品の状態
There are 9 isoforms produced by alternative splicing. -
別名
- AI413597 antibody
- AW045860 antibody
- DDPAC antibody
see all
画像
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All lanes : Anti-Tau (phospho S396) antibody [E178] (ab32057) 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: 60 secondsBlocking/Diluting buffer and concentration 5% NFDM/TBST
Tau assembles into oligomers as described in PMID: 28382304, 32692785 and 30120733.
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Immunohistochemistry analysis of paraffin-embedded human cerebrum tissue sections labeling Tau (phospho S396) with ab32057 at 1/4000 dilution (0.026 μg/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 human cerebrum without alkaline phosphatase treatment (image A). No staining on human cerebrum with alkaline phosphatase treatment (image B).
The section was incubated with ab32057 overnight at +4°C. -
Immunohistochemistry analysis of frozen mouse cerebrum tissue sections labeling Tau (phospho S396) with ab32057 at 1/100 (1μg/mL). ab150077 AlexaFluor®488 Goat anti-Rabbit at 1/1000 (2 μg/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.
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ab32057 (purified) at 1/20 dilution (0.5ug) immunoprecipitating Tau in Human fetal brain lysates.
Lane 1: Human fetal brain lysates 10ug
Lane 2 (+): ab32057 & Human fetal brain lysates
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32057 in Human fetal brain lysates
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST. -
Dot blot analysis of Tau (phospho S396) phospho peptide (Lane 1) and Tau non-phospho peptide (Lane 2) labelling Tau (phospho S396) with ab32057 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.
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Indirect ELISA antigen dose-response curve using ab32057 at 1000-0 ng/mL. Antigen Human Tau (phospho S396) peptide, Human Tau non-phospho peptide at concentration of 1000 ng/mL. Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG H+L at 1/2500 dilution was used as the secondary antibody.
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All lanes : Anti-Tau (phospho S396) antibody [E178] (ab32057) 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 secondsBlocking/Diluting buffer and concentration 5% NFDM/TBST
Tau assembles into oligomers as described in PMID: 28382304, 32692785 and 30120733.
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Immunohistochemistry analysis of paraffin-embedded mouse colon tissue sections labeling Tau (phospho S396) with ab32057 at 1/4000 dilution (0.026 μg/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 ab32057 overnight at +4°C. -
Immunohistochemistry analysis of frozen rat cerebrum tissue sections labeling Tau (phospho S396) with ab32057 at 1/100 (1 μg/mL). ab150077 AlexaFluor®488 Goat anti-Rabbit at 1/1000 (2 μg/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 rat cerebrum, the signal decreased after phosphatase treatment at 37℃ for 2h.
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Immunohistochemistry analysis of paraffin-embedded rat colon tissue sections labeling Tau (phospho S396) with ab32057 at 1/4000 dilution (0.026 μg/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 ab32057 overnight at +4°C. -
IHC image of Tau 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 ab32057, 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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Biotinylated Human Tau (pS396) [0.05 µg/ml] was loaded to SA biosensor on Fortebio RED96e Machine, then associate with recombinant Anti-Tau (phospho S396) antibody [E178] in serial concentration points [3.33, 1.67, 0.83, 0.42, 0.21 nM/mL] by 2-fold dilution, next to dissociate in blank testing buffer [0.1% BSA in PBST (0.05%Tween-20)]. Calculated signals had already subtracted blank control, curve fitting using 1:1 binding model. Non-phospho Tau protein’s association and dissociation were also showed in graph. KD(M) value of Anti-Tau (phospho S396) antibody [E178] is 2.08E-11
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Immunohistochemistical detection of Tau antibody [E178] (ab32057) on formaldehyde fixed paraffin-embedded rat tongue sectionsAntigen retrieval step: Heat mediated. Buffer Used: Citric acid pH6. Permeabilization: None. Primary antibody incubated at 1/1000 for 2 hours @ 21°C in TBS/BSA/azide. Secondary antibody: anti Rabbit IgG Conjugated to Biotin (1/200). A strong pattern of immunostaining which appears to be mostly localised to nerve fibres and their cell bodies (Islet of Langerhans cells are also positive). In submitted image of central area of tongue coloured arrowheads indicate features: red for nerves cut in cross-section (T/S), each brown dot representing a single axon green for what appears to me to be small nerve fibres wrapping around a partial muscle fibre black for a Ganglion containing seven positive nerve cell bodies. Surrounding these are collagen fibres (C), adipocytes (A) and skeletal/striated muscle fibres in L/S ( M-
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Immunohistochemistical detection of Tau antibody [E178] (ab32057) on formaldehyde-fixed paraffin-embedded human salivary gland sections. Antigen retrieval step: Heat mediated. Buffer Used: Citric acid pH6. Permeabilization: No. Blocking step: 1% BSA for 10 mins @ 21°C. ab32057 incubated at 1/1000 for 2 hours @ 21°C in TBS/BSA/azide. Secondary antibody: anti rabbit IgG conjugated to Biotin (1/200). NB: An interesting pattern of positivity that seems to be supported by the Human Protein Atlas. Coloured arrowheads in the submitted image indicate features: red for positive serous glands, blue for positive intra-lobular collecting ducts, black for negative mucous glands (there is a serous demilune around this acinus), yellow for intralobular collecting ducts, green for nerve tracks in the interlobular areas, blue for positive interlobular collecting ducts. There appears to be a population of positive nuclei but this may b
プロトコール
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (60)
ab32057 は 60 報の論文で使用されています。
- Wang L et al. Lactoferrin modification of berberine nanoliposomes enhances the neuroprotective effects in a mouse model of Alzheimer's disease. Neural Regen Res 18:226-232 (2023). PubMed: 35799547
- Schömig C et al. Hippocampal mTOR Dysregulation and Morphological Changes in Male Rats after Fetal Growth Restriction. Nutrients 14:N/A (2022). PubMed: 35276811
- Yang Y et al. Ginsenoside Rg1 improves Alzheimer's disease by regulating oxidative stress, apoptosis, and neuroinflammation through Wnt/GSK-3β/β-catenin signaling pathway. Chem Biol Drug Des 99:884-896 (2022). PubMed: 35313087
- Zinnen AD et al. Alpha-synuclein and tau are abundantly expressed in the ENS of the human appendix and monkey cecum. PLoS One 17:e0269190 (2022). PubMed: 35687573
- Qu Y et al. The neuroprotection of deproteinized calf blood extractives injection against Alzheimer's disease via regulation of Nrf-2 signaling. Aging (Albany NY) 13:11150-11169 (2021). PubMed: 33819182