Product: Phospho-Tau (Ser396) Antibody
Catalog: AF3148
Description: Rabbit polyclonal antibody to Phospho-Tau (Ser396)
Application: WB IHC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Bovine, Horse, Sheep, Rabbit, Dog, Chicken, Xenopus
Mol.Wt.: 50-80kDa; 79kD(Calculated).
Uniprot: P10636
RRID: AB_2834583

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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Pig(100%), Bovine(100%), Horse(100%), Sheep(100%), Rabbit(100%), Dog(100%), Chicken(100%), Xenopus(100%)
Clonality:
Polyclonal
Specificity:
Phospho-Tau (Ser396) Antibody detects endogenous levels of Tau only when phosphorylated at Serine 396.
RRID:
AB_2834583
Cite Format: Affinity Biosciences Cat# AF3148, RRID:AB_2834583.
Conjugate:
Unconjugated.
Purification:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

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

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
P10636 TAU_HUMAN:

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

Description:
This gene encodes the microtubule-associated protein tau (MAPT) whose transcript undergoes complex, regulated alternative splicing, giving rise to several mRNA species. MAPT transcripts are differentially expressed in the nervous system, depending on stage of neuronal maturation and neuron type.
Sequence:
MAEPRQEFEVMEDHAGTYGLGDRKDQGGYTMHQDQEGDTDAGLKESPLQTPTEDGSEEPGSETSDAKSTPTAEDVTAPLVDEGAPGKQAAAQPHTEIPEGTTAEEAGIGDTPSLEDEAAGHVTQEPESGKVVQEGFLREPGPPGLSHQLMSGMPGAPLLPEGPREATRQPSGTGPEDTEGGRHAPELLKHQLLGDLHQEGPPLKGAGGKERPGSKEEVDEDRDVDESSPQDSPPSKASPAQDGRPPQTAAREATSIPGFPAEGAIPLPVDFLSKVSTEIPASEPDGPSVGRAKGQDAPLEFTFHVEITPNVQKEQAHSEEHLGRAAFPGAPGEGPEARGPSLGEDTKEADLPEPSEKQPAAAPRGKPVSRVPQLKARMVSKSKDGTGSDDKKAKTSTRSSAKTLKNRPCLSPKHPTPGSSDPLIQPSSPAVCPEPPSSPKYVSSVTSRTGSSGAKEMKLKGADGKTKIATPRGAAPPGQKGQANATRIPAKTPPAPKTPPSSGEPPKSGDRSGYSSPGSPGTPGSRSRTPSLPTPPTREPKKVAVVRTPPKSPSSAKSRLQTAPVPMPDLKNVKSKIGSTENLKHQPGGGKVQIINKKLDLSNVQSKCGSKDNIKHVPGGGSVQIVYKPVDLSKVTSKCGSLGNIHHKPGGGQVEVKSEKLDFKDRVQSKIGSLDNITHVPGGGNKKIETHKLTFRENAKAKTDHGAEIVYKSPVVSGDTSPRHLSNVSSTGSIDMVDSPQLATLADEVSASLAKQGL

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Horse
100
Bovine
100
Sheep
100
Dog
100
Xenopus
100
Chicken
100
Rabbit
100
Zebrafish
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P10636 As Substrate

Site PTM Type Enzyme
A2 Acetylation
Y18 Phosphorylation P06241 (FYN) , P43405 (SYK)
T39 Phosphorylation
S46 Phosphorylation
T50 Phosphorylation
S56 Phosphorylation
S61 Phosphorylation
S171 Phosphorylation
T173 Phosphorylation
S214 Phosphorylation O00141 (SGK1)
S232 Phosphorylation
S235 Phosphorylation Q00535 (CDK5)
S238 Phosphorylation
S255 Phosphorylation Q13131 (PRKAA1)
S355 Phosphorylation Q13131 (PRKAA1)
S369 Phosphorylation Q13627 (DYRK1A)
T386 Phosphorylation
S388 Phosphorylation
S396 Phosphorylation P49841 (GSK3B) , Q13131 (PRKAA1) , P48730 (CSNK1D)
S400 Phosphorylation P49841 (GSK3B)
S411 Phosphorylation
T449 Phosphorylation
S451 Phosphorylation
T466 Phosphorylation
T470 Phosphorylation
T492 Phosphorylation
T498 Phosphorylation P53779 (MAPK10) , P45983 (MAPK8) , P49841 (GSK3B) , Q00535 (CDK5) , P45984 (MAPK9)
S501 Phosphorylation
S502 Phosphorylation
S508 Phosphorylation
S512 Phosphorylation P49841 (GSK3B)
Y514 Phosphorylation
S515 Phosphorylation P49841 (GSK3B) , Q5TCY1 (TTBK1)
S516 Phosphorylation Q00535 (CDK5) , Q5TCY1 (TTBK1) , P45984 (MAPK9) , P53779 (MAPK10) , P49841 (GSK3B) , P45983 (MAPK8)
S519 Phosphorylation Q13627 (DYRK1A) , P53779 (MAPK10) , P45983 (MAPK8) , P48730 (CSNK1D) , Q5TCY1 (TTBK1) , Q00535 (CDK5) , P49841 (GSK3B) , P45984 (MAPK9) , P28482 (MAPK1)
T522 Phosphorylation P45983 (MAPK8) , P48730 (CSNK1D) , P49841 (GSK3B) , Q00535 (CDK5) , P28482 (MAPK1) , P53779 (MAPK10)
S525 O-Glycosylation
S525 Phosphorylation
S527 Phosphorylation
T529 Phosphorylation P45984 (MAPK9) , Q8IWQ3 (BRSK2) , P45983 (MAPK8) , P49841 (GSK3B) , Q8TDC3 (BRSK1) , Q13627 (DYRK1A) , P53779 (MAPK10) , Q92630 (DYRK2) , P57059 (SIK1) , Q00535 (CDK5)
S531 Phosphorylation P49841 (GSK3B) , P57059 (SIK1) , P17612 (PRKACA) , Q00535 (CDK5)
T534 Phosphorylation Q00535 (CDK5) , P53779 (MAPK10) , P49841 (GSK3B) , P45983 (MAPK8)
T537 Phosphorylation
T548 Phosphorylation P45984 (MAPK9) , Q9UQM7 (CAMK2A) , P17612 (PRKACA) , P49841 (GSK3B) , Q00535 (CDK5)
S552 Phosphorylation Q9UQM7 (CAMK2A) , Q00535 (CDK5)
S554 Phosphorylation
S555 O-Glycosylation
S555 Phosphorylation
S558 Phosphorylation
K571 Ubiquitination
S575 Phosphorylation
S579 Phosphorylation P57059 (SIK1) , Q9P0L2 (MARK1) , P17612 (PRKACA) , Q8TDC3 (BRSK1) , P49841 (GSK3B) , P49840 (GSK3A) , Q9UQM7 (CAMK2A) , O96017 (CHEK2) , Q8IWQ3 (BRSK2)
T580 Phosphorylation
S602 Phosphorylation
S610 Phosphorylation P57059 (SIK1)
K611 Methylation
S622 Phosphorylation P57059 (SIK1)
Y627 Phosphorylation
K628 Acetylation
K628 Ubiquitination
S633 Phosphorylation
T636 Phosphorylation
S637 Phosphorylation Q16512 (PKN1)
S641 Phosphorylation P49840 (GSK3A) , Q9P0L2 (MARK1) , P57059 (SIK1)
S669 Phosphorylation
K670 Ubiquitination
S673 Phosphorylation Q9P0L2 (MARK1) , P17612 (PRKACA) , P57059 (SIK1)
K686 Acetylation
K687 Acetylation
T690 Phosphorylation
T694 Phosphorylation
Y711 Phosphorylation
S713 Phosphorylation Q00535 (CDK5) , P49840 (GSK3A) , P45984 (MAPK9) , P45983 (MAPK8) , P53779 (MAPK10) , P49841 (GSK3B)
S717 O-Glycosylation
S717 Phosphorylation P49841 (GSK3B)
T720 Phosphorylation
S721 Phosphorylation P45984 (MAPK9) , P49841 (GSK3B) , Q00535 (CDK5) , P45983 (MAPK8) , Q13627 (DYRK1A) , P28482 (MAPK1) , P53779 (MAPK10) , P49840 (GSK3A) , P48730 (CSNK1D)
S726 Phosphorylation P17612 (PRKACA)
S729 Phosphorylation
S730 Phosphorylation
T731 Phosphorylation
S733 Phosphorylation
S739 Phosphorylation P53779 (MAPK10) , P45984 (MAPK9) , P45983 (MAPK8) , Q5TCY1 (TTBK1)
T744 Phosphorylation

Research Backgrounds

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/MAPT 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.

PTMs:

Phosphorylation at serine and threonine residues in S-P or T-P motifs by proline-directed protein kinases (PDPK1, CDK1, CDK5, GSK3, MAPK) (only 2-3 sites per protein in interphase, seven-fold increase in mitosis, and in the form associated with paired helical filaments (PHF-tau)), and at serine residues in K-X-G-S motifs by MAP/microtubule affinity-regulating kinase (MARK1, MARK2, MARK3 or MARK4), causing detachment from microtubules, and their disassembly. Phosphorylation decreases with age. Phosphorylation within tau/MAP's repeat domain or in flanking regions seems to reduce tau/MAP'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. Phosphorylation at Ser-548 by GSK3B reduces ability to bind and stabilize microtubules. Phosphorylation at Ser-579 by BRSK1 and BRSK2 in neurons affects ability to bind microtubules and plays a role in neuron polarization. Phosphorylated at Ser-554, Ser-579, Ser-602, Ser-606 and Ser-669 by PHK. Phosphorylation at Ser-214 by SGK1 mediates microtubule depolymerization and neurite formation in hippocampal neurons. There is a reciprocal down-regulation of phosphorylation and O-GlcNAcylation. Phosphorylation on Ser-717 completely abolishes the O-GlcNAcylation on this site, while phosphorylation on Ser-713 and Ser-721 reduces glycosylation by a factor of 2 and 4 respectively. Phosphorylation on Ser-721 is reduced by about 41.5% by GlcNAcylation on Ser-717. Dephosphorylated at several serine and threonine residues by the serine/threonine phosphatase PPP5C.

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.

O-glycosylated. O-GlcNAcylation content is around 8.2%. There is reciprocal down-regulation of phosphorylation and O-GlcNAcylation. Phosphorylation on Ser-717 completely abolishes the O-GlcNAcylation on this site, while phosphorylation on Ser-713 and Ser-721 reduces O-GlcNAcylation by a factor of 2 and 4 respectively. O-GlcNAcylation on Ser-717 decreases the phosphorylation on Ser-721 by about 41.5%.

Glycation of PHF-tau, but not normal brain TAU/MAPT. 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.

Subcellular Location:

Cytoplasm>Cytosol. Cell membrane>Peripheral membrane protein>Cytoplasmic side. Cytoplasm>Cytoskeleton. Cell projection>Axon. Cell projection>Dendrite.
Note: Mostly found in the axons of neurons, in the cytosol and in association with plasma membrane components.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
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.

Subunit Structure:

Interacts with MARK1, MARK2, MARK3 AND MARK4. Interacts with PSMC2 through SQSTM1 (By similarity). Interacts with SQSTM1 when polyubiquitinated. Interacts with FKBP4 (By similarity). Binds to CSNK1D. Interacts with SGK1. Interacts with EPM2A; the interaction dephosphorylates MAPT at Ser-396. Interacts with PIN1. Interacts with LRRK2.

Family&Domains:

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

Research Fields

· Environmental Information Processing > Signal transduction > MAPK signaling pathway.   (View pathway)

· Human Diseases > Neurodegenerative diseases > Alzheimer's disease.

References

1). Andrographolide Protects PC12 Cells Against β-Amyloid-Induced Autophagy-Associated Cell Death Through Activation of the Nrf2-Mediated p62 Signaling Pathway. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, 2018 (PubMed: 30235892) [IF=5.6]

Application: WB    Species:    Sample: PC12 cells

Figure 9. |Effect of Nrf2 siRNA on p62, p21, and p-tau/tau protein expression levels in PC12 cells. (C,D) After transfection with 20 µM of Nrf2 siRNA, cells were then pre-treated with Andro (20 µM) for 1 h followed by stimulation with Aβ (10 µM) for an additional 24 h. Then, p62, p21, and p-tau/tau protein expression levels were evaluated by Western blot analysis. * p < 0.05 versus the blank control;# p < 0.05 versus Andro+Aβ1–42 group were considered statistically significant differences.

2). Ginsenoside Rg1 attenuates LPS-induced cognitive impairments and neuroinflammation by inhibiting NOX2 and Ca2+–CN–NFAT1 signaling in mice. Journal of Functional Foods, 2021 [IF=5.6]

Application: WB    Species: Mouse    Sample: brain tissue

Fig. 3. Effects of Rg1 on the expressions of MAP2, PSD95, Tau, p-Tau, Bax and Bcl-2 in brain tissue of LPS-exposed mice (immunoblot). (A) The bands and relative expression of MAP2 over control. (B) The bands and relative expression of p22phox over control. (C) The bands and relative expression of Tau over control. (D) The bands and relative expression of p-Tau/Tau over control. (E) The bands and relative expression of Bax over control. (F) The bands and relative expression of Bcl-2 over control. Results are expressed as the mean ± SD, n = 4. **P < 0.01 compared with control group; #P < 0.05, ##P < 0.01 compared with LPS group.

3). Baicalin Attenuates Panton-Valentine Leukocidin (PVL)-Induced Cytoskeleton Rearrangement via Regulating the RhoA/ROCK/LIMK and PI3K/AKT/GSK-3β Pathways in Bovine Mammary Epithelial Cells. International journal of molecular sciences, 2023 (PubMed: 37833969) [IF=5.6]

4). Maresin 1 alleviates sevoflurane-induced neuroinflammation in neonatal rats via JAK2/STAT3/IL-6 pathways. International Immunopharmacology, 2022 (PubMed: 35729840) [IF=5.6]

Application: WB    Species: Rat    Sample: hippocampus

Fig. 2. MaR1 decreased sevoflurane-induced hyperphosphorylation level of Tau at multi-sites. A-F Rats’ hippocampi were collected and analyzed by Western blotting using phosphorylated Tau antibodies (A) and quantitative analysis (B-F). One-way ANOVA, [pT205] F(3, 8) = 7.321, p = 0.0123; [pS214] F(3, 8) = 5.542, p = 0.0315; [pS262] F(3, 8) = 5.653, p = 0.0373; [pS396] F(3, 8) = 8.763, p = 0.0153; [pS404] F(3, 8) = 7.321, p = 0.0184. n = 3. * p < 0.05; # p < 0.05. Data were presented as mean ± SEM.

5). Therapeutic effects of total saikosaponins from Radix bupleuri against Alzheimer’s disease. Frontiers in Pharmacology, 2022 (PubMed: 35935875) [IF=5.6]

Application: WB    Species: Mice    Sample: brain

FIGURE 5 Effects of TS on expressions of p-tau, NDP52, p62 and LC3-II proteins in APP/PS1 mice. (A) The effect of TS on p-tau (ser396) in hippocampus and cortex of APP/PS1 mice was investigated by immunohistochemistry. (B) Quantitative analysis of the percentage of p-tau (Ser396) positive area in the hippocampus of APP/PS1 mice (#### p < 0.0001, ***p = 0.0007, ****p < 0.0001). (C) Quantitative analysis of the percentage of p-tau (Ser396) positive area in the cortex of APP/PS1 mice (#### p < 0.0001, **p = 0.0016, ****p < 0.0001). (D) The effects of TS on expressions of p-tau (Ser 396), NDP52, p62 and LC3-II proteins in the brain of APP/PS1 mice were detected by Western blot. (E) Quantitative analysis of the relative expression level of p-tau (Ser 396) protein in each group (# p = 0.0332, * p = 0.0436, ** p = 0.0042). (F) Quantitative analysis of the relative expression level of NDP52 protein in each group (### p = 0.0008, *p = 0.0177, **p = 0.0086). (G) Quantitative analysis of the relative expression level of p62 protein in each group (# p = 0.0379, **p = 0.0086). (H) Quantitative analysis of the relative expression level of LC3-II protein in each group (# p = 0.0216, ** p = 0.0073). (I) Quantitative analysis of the relative expression level of Beclin-1 protein in each group (# p = 0.0496, *p = 0.0136). #Compared with WT group; *Compared with model group. Data are presented as mean ± SEM (n = 3).

6). Cerebroprotein hydrolysate attenuates neurodegenerative changes in Alzheimer’s mice model via ferroptosis pathway. Frontiers in Pharmacology, 2023 [IF=5.6]

7). MiR-29a efficiently suppresses the generation of reactive oxygen species and α-synuclein in a cellular model of Parkinson's disease by potentially targeting GSK-3β. European journal of pharmacology, 2024 (PubMed: 38685306) [IF=5.0]

8). Carnitine palmitoyltransferase 1 (CPT1) alleviates oxidative stress and apoptosis of hippocampal neuron in response to beta-Amyloid peptide fragment Aβ25-35. Bioengineered, 2021 (PubMed: 34424821) [IF=4.9]

Application: WB    Species: Mice    Sample: Aβ25-35-induced HT22 cells

Figure 5. CPT1C overexpression decreased the deposition of AD marker proteins in Aβ25-35-induced HT22 cells. Following transfection of Ov-CPT1C or Ov-NC for 24 h, HT22 cells were treated with Aβ25–35 for another 24 h (a) The mRNA expressions of App, p-Tau and Bace-1 were evaluated using RT-qPCR. (b) The protein expressions of App, p-Tau and Bace-1 were evaluated using western blot. ***P < 0.001 vs. Control group, ###P < 0.001 vs Aβ25-35 + Ov-NC.

9). Long-Term Administration of Triterpenoids From Ganoderma lucidum Mitigates Age-Associated Brain Physiological Decline via Regulating Sphingolipid Metabolism and Enhancing Autophagy in Mice. Frontiers in Aging Neuroscience, 2021 (PubMed: 34025387) [IF=4.8]

Application: WB    Species: mouse    Sample: brain

FIGURE 9 | Ameliorate effects of Ganodenic acid A by regulating sphingolipid metabolism in 3 × Tg-AD mice. (A) Experimental procedure in 3 × Tg-AD mice;(B) The AD biomarkers of p-Tau, Aβ, APOE, TREM2, CD33, the inflammatory cytokines of TNF-α and NF-κB p65 and the autophagy level of LC3A/B were measured in brain tissues

10). Regular Exercise Enhances Cognitive Function and Intracephalic GLUT Expression in Alzheimer\'s Disease Model Mice. JOURNAL OF ALZHEIMERS DISEASE, 2019 (PubMed: 31561359) [IF=4.0]

Application: WB    Species: mouse    Sample: brain

Fig. 1. The expression A and P-Tau. Representative immunoblots of A and P-Tau in the cortex (A-C) and hippocampus (D-F) in each group. Protein immunoreactivity was normalized to -actin. Individual data are presented as the mean ± S.E.M. from four individual mice in each group.

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