44-750G

antibody from Invitrogen Antibodies

Targeting: MAPT DDPAC, FLJ31424, FTDP-17, MAPTL, MGC138549, MSTD, MTBT1, MTBT2, PPND, PPP1R103, tau

Provider product page for 44-750G

Western blot

Immunohistochemistry

Flow cytometry

Other assay

Antibody data

Antibody Data
Antigen structure
References [51]
Comments [0]
Validations
Western blot [2]
Immunohistochemistry [2]
Flow cytometry [1]
Other assay [33]

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Product number: 44-750G - Provider product page
Provider: Invitrogen Antibodies
Product name: Phospho-Tau (Ser262) Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Synthetic peptide
Description: The antibody has been negatively preadsorbed using a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated Tau. The final product is generated by affinity chromatography using a Tau-derived peptide that is phosphorylated at serine 262. Positive control used: Recombinant human Tau +/- PKA, or African green monkey kidney (CV-1) cells, stably expressing human four repeat Tau and SV40 small T antigen to induce specific inhibition of PP2A.
Reactivity: Human, Mouse, Rat
Host: Rabbit
Isotype: IgG
Vial size: 100 µL
Storage: -20°C

MAPT protein structure - 44-750G shown in red.

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Experimental details: Immunohistochemistry analysis of Phospho-Tau (pS262) showing staining in the cytoplasm of paraffin-embedded human astroglioma tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Phospho-Tau (pS262) polyclonal antibody (Product # 44-750G) diluted in 3% BSA-PBS at a dilution of 1:100 overnight at 4ºC in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

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Experimental details: Immunohistochemistry analysis of Phospho-Tau (pS262) showing staining in the cytoplasm of paraffin-embedded mouse brain tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Phospho-Tau (pS262) polyclonal antibody (Product # 44-750G) diluted in 3% BSA-PBS at a dilution of 1:20 overnight at 4ºC in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

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Experimental details: Figure 6 Screen for additional pJNK-Tau sites in striatum. (A) Phosphorylation of Tau protein at epitopes subject to phosphorylation by JNK was analyzed by immunoblot. (B) Band optical density (OD) from phosphorylation-specific probes relative to total Tau expression is presented as percent of 2 month-old WT (mean +- SEM) and was analyzed by two-way ANOVA with Bonferroni post-hoc tests comparing each A53T group to age-matched controls (*p

Supportive validation

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Experimental details: Figure 7 Analysis of tau signal with polyclonal antibodies by Western blotting. Proteins were extracted from the cortex of 3 mouse lines: control mice (WT and Hypothermic), Tau KO mice and 3xTg-AD mice. Proteins were separated by SDS-PAGE and then identified with the following polyclonal antibodies: A: Total Tau, B: pS199, C: pS396, D: pS404, E: pT205, F: pS422, G: pS262 and H: pS409. Normal anti-rabbit secondary antibodies were used to detect primary antibodies. The heat stable fraction was used to remove non-specificity: I: pS262 and J: pS409. Quantifications of the blots are available in Figure S5 .

Supportive validation

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Experimental details: Figure 6 Neuronal uptake of PBS-soluble HMW tau derived from human AD brain. ( a , b ) Primary neurons were incubated with AD or control brain extracts (cases were matched for age and postmortem interval ( Supplementary Table S1 )) and immunostained at day 2 ( a ). ( b ) Quantification of fluorescence intensity of human tau staining. One-way ANOVA and a subsequent Tukey-Kramer test. ( c , d ) Tau uptake ( c ) and seeding activity ( d ) assay in HEK-tau-biosensor cells. (Mann-Whitney U -test) ( e ) Subcellular localization of human tau taken up by neurons (PBS-3,000 g , 500 ng ml -1 human tau). ( f ) Neuron-to-neuron transfer of tau in a 3-chamber microfluidic device. AD brain extract (PBS-3,000 g , 500 ng ml -1 human tau) was added to the 1st chamber. Human tau positive neurons were detected in both the 1st and 2nd chamber at day 7 (arrow). ( g , h ) Quantification of total-tau ( g ) and phospho-tau ( h ) levels in AD and control brain extract (ELISA). Unpaired t -test. ( i ) Brain extracts were immunoblotted with phospho-tau specific antibodies recognizing different epitopes. Representative immunoblot and quantification of phospho-tau levels at each epitope. Unpaired t -test. ( j , k ) SEC analysis of PBS-soluble tau from AD and control brain. ( j ) Representative graph of total tau levels (ELISA) in SEC-separated samples. Small peaks for HMW fractions were detected in both groups (right panel). ( k ) Mean total tau levels of HMW SEC fractions. ( l ) Tau uptake from each SEC

Supportive validation

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Experimental details: Figure 5 Higher level of total tau, phospho-tau and cleaved tau in P301S mice receiving an Abeta 1-42 injection into the entorhinal cortex as compared with Abeta 1-40 injection. 8 month old P301S mice were injected with Abeta1-40 into the left entorhinal cortex and Abeta1-42 into the right entorhinal cortex. 18 days after injection, the brain extracts were analyzed by western blot. A) Representative images of western blots of soluble tau (S1 fraction) detected with anti-tau antibodies and normalized to actin. B) Quantification of western blots in (A) . C) Representative images of western blots of insoluble tau (P1 fraction) detected with anti-tau antibodies. Both S1 brain fraction and P1 brain fraction show a significant increase in HT7, PHF-1 and pS262 signal in entorhinal cortex injected with Abeta1-42 compared with Abeta1-40 injected entorhinal cortex. Also more cleaved tau (Tau421) was detected in the S1 fraction from the Abeta1-42 injected brain. D) Quantification of western blots in (C) . Histographs show mean +/- standard error, n =3. *p

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Experimental details: Figure 6 Abeta 1-42 treatment increases pathological tau recognized by AT180 when compared with Abeta 1-40 treatment. Brain sections were immunostained with AT180, which recognizes p-tau at Thr 231 . (A) Representative images for pathological tau detected with AT180. (B) Higher magnification showed significantly increased tau tangles in Abeta1-42 injected entorhinal cortex (C) and hippocampus (B) . Magnification: 4x for A; 10x for B and C. Scale bar: 100um.

Supportive validation

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Experimental details: Figure 7 Abeta 1-42 promotes tau phosphorylation, cleavage and aggregation while Abeta 1-40 does not. 8 month old P301S mice (1N4R form with Prion promoter) were injected with saline into the left entorhinal cortex and Abeta1-42 (2ug) into the right entorhinal cortex or saline into the left entorhinal cortex and Abeta1-40 (2ug) into the right entorhinal cortex. 18 days after injection, the brain extracts were analyzed by western blot. A) Representative images of western blots of soluble tau (S1 fraction) detected with anti-tau antibodies and normalized to actin. B) Quantification of western blots in (A) . C) Representative images of western blots of insoluble tau (P1 fraction), detected with anti-tau antibodies. D) Quantification of western blots in (C) Abeta1-42 injection significantly induced tau phosphorylation (PHF-1 and pS262 epitope) in the S1 and P1 brain fractions. 3-fold increase in Tau421 was found in the soluble fraction (A and B) . Significant decrease in pS262 immunoreactivity was observed in S1 fraction from Abeta1-40 injected brain areas compared to control side. Histographs show mean +/- standard error, n =3. *p

Supportive validation

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Experimental details: Figure 3 Abeta1-42 monomers promote phosphorylation at particular sites that have been related to Alzheimer's disease (AD) progression. Representative Western blot of brain extracts (20 mug protein) from control (saline) and treated (Abeta1-42 peptides by ICV for 3 h) mice using antibodies specific for the detection four pathological Tau phosphorylation sites: AT8, pS396, pS262, and pS422. An antibody raised against GAPDH or Tau 5 served as loading control. Densitometric quantification shows an increase of the total protein level of AT8, pS396, and pS422 induced by monomers while monomeric and oligomeric preparations did not change pS262 expression. The data are mean +- standard error of the mean (SEM), * P < 0.05; *** P < 0.001 vs. control by one-way ANOVA followed by Bonferroni post hoc test, n = 6.

Supportive validation

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Experimental details: Figure 3 MB reduced Tau phosphorylation by overexpressed MARK4 in 293T cells. ( a ) Representative immunoblot of lysates from 293T cells transfected with 4R2N Tau alone, HA-MARK4 (KD and WT) alone or Tau + HA-MARK4. Cells transfected with Tau + HA-MARK4-WT were treated with AC, MB and OS for 3 h at concentrations indicated in the panel. The levels of pTau S262 and total Tau were reported with beta-actin as the loading control. ( b ) Quantification of the ratio of monomeric pTau S262 to monomeric total Tau presented as the percent of the level found in cells transfected with Tau and MARK4-WT but without drug treatment. Results are mean +- SD (n = 3). *0.01 < P < 0.05; ***P < 0.001. Student's t test is conducted between corresponding values in AC and OS or MB and OS group. ( c ) Representative immunoblot of lysates from 293T cells transfected with Tau + HA-MARK4-KD followed by treatment with AC, MB and OS as described in A). beta-actin blots were cropped. Full blots are shown in Supplementary Fig. S4 .

Supportive validation

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Experimental details: Figure 4 MB reduced Tau phosphorylation by endogenous kinases in 293T cells. ( a ) Representative immunoblot of lysates from 293T cells transfected with 4R2N Tau. Transfected cells were treated with AC, MB and OS for 3 h at concentrations indicated in the panel. The levels of pTau S262, S396, S214, S202 and total Tau were reported with beta-actin as the loading control. ( b-e ) Quantification of the ratio of pTau to total Tau presented as the percent of the level found in cells transfected with Tau but without drug treatment. Results are mean +- SD (n = 3). *0.01 < P < 0.05; **0.001 < P < 0.01 ***P < 0.001. Student's t test is conducted between corresponding values in AC and OS or MB and OS group.

Supportive validation

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Experimental details: Figure 6 MB directly inhibits the ability of MARK4 to phosphorylate Tau in an in vitro kinase assay. GST-MARK4 was incubated with purified His-Tau and ATP in the presence of increasing concentration of AC, MB or OS for 30 min at 30 degC. ( a ) Representative immunoblot of the kinase reaction probed by antibodies against GST, pTau S262 and total Tau. ( b ) Quantification of the ratio of monomeric pTau S262 to monomeric total Tau presented as the percent of the level without drug treatment. Results are mean +- SD (n = 4). *0.01 < P < 0.05; **0.001 < P < 0.01. Student's t test is conducted between corresponding values in AC and OS or MB and OS group.

Supportive validation

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Experimental details: Figure 8. Inhibition of MT dynamics causes tau hyperphosphorylation and spine loss though tau. (A and B) WB analysis of Glu and acetylated (Acety) tubulin (A) as well as phosphorylated (AT8 and pS262) and total (tau-46) tau levels (B) in hippocampal neurons (7 DIV) incubated with 10 nM taxol for 3 h. (C) Quantification of normalized AT8 and pS262 against control levels in taxol-treated neurons. (D) WB analysis of total tau (tau-C) and tubulin (DM1A) levels in cortical neurons (7 DIV) treated with 10 nM taxol for 3 h. Aliquots of soluble (S) and MT pellet (P) fractions are shown. (E) Representative images from time-lapse recordings of NIH3T3 cells cotransfected with human 2N4R WT or S262A tau-GFP and RFP-tubulin before and 10 min after 10 nM taxol. Dashed lines indicate cell peripheries. Bar, 20 um. (F) Quantification of background-subtracted MT-bound WT tau, S262A tau, and MT intensity after taxol treatment. (G and H) Representative images (G) and quantification (H) of spine density and morphology of hippocampal neurons (21 DIV) treated with vehicle control, Abeta, or taxol for the indicated times +- washout (w/o). (I and J) Representative images (I) and quantification (J) of spine density and morphology of hippocampal neurons (21 DIV) from WT and tau-KO mice treated with vehicle control, Abeta, or taxol for the indicated times. Bars, 2 um. Gray asterisks in J compare Abeta- or taxol-treated KO neurons to KO control neurons. Data are shown as means +- SEM. *, P < 0.5; **,

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Experimental details: Figure 3 DAU attenuated tau phosphorylation in both N2a/APP cells and HEK293/Tau cells. Levels of phosphorylated tau and total tau in N2a/WT and N2a/APP cells (a, b) and in HEK293/Tau cells (c, d) as determined by Western blot analysis. beta -Actin was used as a loading control. N = 3. Data show the mean +- SEM. * P < 0.05 compared to N2a/WT cells, # P < 0.05 and ## P < 0.01 compared to untreated N2a/APP cells, and & P < 0.05 and &&& P < 0.001 compared to untreated HEK293/Tau cells.

Supportive validation

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Experimental details: Figure 1 Tau acetylation modulates tau phosphorylation at specific epitopes. (a-c ) Immunoblot analysis and quantification using the indicated tau antibodies for (a ) cells expressing WT tau-T40 and either an active form of CBP or a catalytically inactive form of CBP (CBP-LD), ( b ) cells co- expressing tau-T40 (WT, DeltaKK, 4KQ, or 4KR) and tau modifying enzymes, and ( c ) cells expressing tau-T40 (WT, DeltaKK, 2KQ, 4KQ, or 4KR) and treated with control (DMSO) or okadaic acid (OA), where indicated. Solid black arrows highlight the ~75 kDa phospho-tau species that is reduced upon acetylation. Statistical significance was assessed using a student t-test (****p < 0.0001). Cropped images from full size immunoblots are provided in panels a-c. Full-length immunoblots are presented in Supplementary Fig. S1 . ( d ) Immunofluorescence microscopy of primary cortical neurons expressing WT-tau-GFP (left) or 4KQ-tau-GFP (right) detecting phosphorylated tau (AT8). White arrows identify transfected neurons. Scale bar, 50 um.

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Experimental details: Amylin, tau, and Abeta inclusions in pancreatic beta cells. Immunohistochemistry is shown for amylin (A), total tau (B), Thr181 phosphorylated tau (AT270; C), Ser262 phosphorylated tau (D), Ser202-Thr205 phosphorylated tau (AT8; E), Thr212-Ser214 phosphorylated tau (AT100; F), Thr231 phosphorylated tau (AT180; G), Ser422 phosphorylated tau (H), Asp421 cleaved tau (I), tau Alz50 (J), tau MC-1 (K), endocrine oligomeric tau (L), exocrine oligomeric tau (M), and Abeta inclusions (N) in pancreatic cells from a 75-year-old male with Alzheimer disease and no history of type 2 diabetes mellitus. Magnification = x40, scale bars = 50mum. The boxed areas (H-N) were examined at a higher magnification: x63 magnification, scale bars = 20mum.

Supportive validation

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Experimental details: Fig. 3 Tau and Abeta inclusions in pancreatic beta cells of subjects with Parkinson''s disease, Lewy body dementia and incidental Lewy body disease. Immunohistochemistry for Ser202-Thr205-phosphorylated tau (AT8) ( a ), Ser262-phosphorylated tau ( b ), Thr181-phosphorylated tau (AT270) ( c ), Thr212-Ser214-phosphorylated tau (AT100) ( d ), Thr231-phosphorylated tau (AT180) ( e ), Ser422-phosphorylated tau ( f ), Asp421-cleaved tau ( g ), tau Alz50 ( h ), tau MC-1 ( i ), endocrine oligomeric tau ( j ), exocrine oligomeric tau ( k ) and Abeta ( l ) inclusions in pancreatic cells from subjects with incidental Lewy body disease ( a , b , c , f ), Parkinson''s disease ( d , e , g , h , i , l ) and dementia with Lewy body ( j , k ): a 40 x magnification; scale bar = 50 um

Supportive validation

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Experimental details: Figure 6 Hippocampal and cortical phosphorylated tau is elevated after single (sTBI) or repeated TBI (rTBI). (A-C) Representative confocal images (63x, multi-plane) show immunolabeling of phosphorylated tau (pTau S262, green) within the CA1 region of the hippocampus (left) and Layer V of the cortex (right) ipsilateral to injury in sham (A) , sTBI (B) , and rTBI (C) animals. (D) Bar graphs show a significant increase in hippocampal phosphorylated tau burden in the rTBI group (black) relative to the sTBI (gray) and sham groups (white; n = 16 slices/4 rats for each treatment condition). (E) Bar graphs show a significant increase in cortical phosphorylated tau burden in the rTBI group (black) relative to the sTBI (gray) and sham groups (white; n = 16 slices/4 rats for each treatment condition). Representative images were obtained on a Leica Microsystems SP8 confocal microscope. The injury was over the right sensorimotor cortex. * p

Supportive validation

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Experimental details: Figure 2 Effects of loganin on AD pathology and the expression of Abeta-related key proteins. ( A ) The relative expression of Abeta deposition. ( B , C ) The relative expression of APP expression and processing related proteins. ( D ) The relative expression of Tau phosphorylation. Scale bar = 100 mum. Data were expressed as mean +- SEM, * p < 0.05, *** p < 0.001, n = 3 for each group.

Supportive validation

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Experimental details: Amyloid-beta (Abeta) aggravates tau protein hyperphosphorylation and accumulation. (A) Western blot analysis of tau phosphorylation using phosphorylation site-specific antibodies. Protein extracts were prepared from 25 adult fly heads. ELAV-Gal4 was used to drive Tau* and Abeta* expression in the fly central nervous system (CNS). Tubulin was used as a loading control. (B) The quantitative results for panel A. The pS262, PHF-1, and AT8 levels were normalized to the Tau-5 antibody signal. (C) Western blot analysis of tau hyperphosphorylation in HEK293 cells with Tau*-RFP expression. Tau*-RFP cells were treated with 10 muM Abeta 40 peptide for 24 hours. Tubulin was used as the loading control. (D) The quantitative results for panel C. Western blot results were calculated using ImageJ software. Levels of pS262 and PHF-1 were normalized to tau5. (E) Western blot analysis of the levels of soluble and insoluble tau protein. Tubulin was used as the loading control. Protein extracts were prepared from 50 adult fly heads. ELAV-Gal4 was used to drive Tau* and Abeta* expression in the fly CNS. (F) The quantitative results for panel E. The tau protein level was normalized to tubulin. Data are presented as the mean +- SEM. * P < 0.05, ** P < 0.01, *** P < 0.001 (unpaired t -test). All western blot analyses were performed three times, and representative blots are shown.

Supportive validation

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Experimental details: Figure 3 A beta1-42 promotes phosphorylation, cleavage and aggregation of tau by increasing GSK3beta activity and by activating pro-caspase3. Sequential extraction was performed on Tau441-YFP-transfected cells treated with transfection reagent alone (control), 200 nM Abeta1-40 or Abeta1-42 for 24 h. Triton fraction was obtained by extracting intracellular tau with 1% Triton. Triton-insoluble fraction was then solubilized with 1% SDS to get SDS fraction. A) Triton fraction was electrophoresed and immunoblotted for total tau (HT7), p-tau(Ser 396/404 ) (PHF-1), p-tau(Thr 262 ) (pS262), Tau421(tau-C3) and beta-actin. beta-Actin was measured from the same blots to ensure that equal protein was present in every lane. Abeta1-42 significantly induced p-tau at Thr 231 and increased levels of Tau421 were observed in Abeta1-42 treated cells, while no change occurred in Abeta1-40 treated cells when compared to cells without treatment. B) Quantification of western blots in (A) . C) Representative images of western blots for SDS fraction with HT7, PHF-1 and pS262. All 3 forms of tau increased in Abeta1-42 treated cells. No change in tau level was found in Abeta1-40 treated cells. There was no detectable signal for tau-C3 in SDS fraction. D) Quantification of western blots in (C) . E) Triton fraction was also blotted with anti-GSK3beta antibody, anti-p-GSK3beta (Y216) and anti-caspase-3 antibody. Abeta1-42 treatment resulted in an increase in phosphorylation of GSK3beta, which represents hi

Supportive validation

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Experimental details: Fig. 1 Tau phosphorylation increases at multiple sites in the brain following r-mTBI in 3xTg-AD mice. Adult female 3xTg-AD mice, 4-- months of age, were subjected to repetitive mild closed-head brain impacts, totaling five with 48-h inter-impact intervals, and sacrificed 24 h after the final impact. A) Representative western blots showing the level of indicated proteins/phosphorylation sites. B) Densitometric quantification of the blots. Among the phosphorylation sites examined, tau was hyperphosphorylated at pThr 205 , pSer 262 and PHF-1 (pSer 396/404 ) sites. Data are presented as scatter dot plots with mean+-SEM ( n = 6-7 mice each) and analyzed using unpaired t test, with Welch's correction in the case of unequal variance.

Supportive validation

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Experimental details: Fig. 5 Translocated I 2 PP2A /SET is colocalized with hyperphosphorylated tau in the neuronal cytoplasm in cerebral cortex of 3xTg-AD mice with r-mTBI. Brain sections were fluorescently dual immuno-stained for I 2 PP2A and phospho-tau. Translocated I 2 PP2A colocalized with pThr 205 -tau, pSer 396 -tau and with pSer 262 -tau in the cytoplasm of neurons (arrows). However, there was no somatodendritic phospho-tau staining in neurons where cytoplasmic I 2 PP2A was absent (filled arrowheads). Scale bar = 10 mu m for all images.

Supportive validation

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Experimental details: Figure 4 Western blot of Tau46 (total Tau) and phosphorylated Tau by GSK3beta and PKA kinases from cerebrospinal fluid (CSF) sampling of four groups: normal, Alzheimer's disease (AD), neurological disorders (NAD), and mild cognitive impairment (MCI). ( A ) Western blotting with Tau 46, Tau Ser205, Tau Ser231, and Tau Ser396 by GSK3beta or Tau Ser214, Tau Ser262, and Tau Ser409 by PKA antibodies. Albumin was used as an internal control. ( B ) Statistical analysis. Bar graphs represent the mean +- SD of triplicates. * p < 0.05, ** p < 0.01, *** p < 0.001 compared with the control group.

Supportive validation

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Experimental details: Figure 2 Lack of PBS-soluble phosphorylated HMW tau species is associated with low tau uptake in primary neurons. ( a , top) Uptake of human tau from brain extracts from rTg4510 and rTg21221 mice by primary neurons (PBS-3,000 g , 500 ng ml -1 human tau). Neurons were immunostained with human tau-specific antibody (green) and total (human and mouse) tau antibody (red). ( a , bottom) Tau uptake assay in HEK-tau-biosensor cells. Brain extracts (10 mug protein) were applied to the cells (lipofectamie (-)). ( n =4) Unpaired t -test. Scale bar, 50 mum. ( b ) Human tau levels in brain extracts (ELISA). ( c ) Immunoblot analysis of PBS-soluble extracts with total tau antibody (DA9). Up-shifted bands in rTg4510 brain suggest phosphorylation of tau (arrow). ( d ) Brain extracts were immunoblotted with phospho-tau specific antibodies recognizing different epitopes. Representative immunoblot and quantification of phospho-tau levels at each epitope. ( n =3-4) Unpaired t -test. ( e , f ) SEC analysis of PBS-soluble tau. ( e ) Representative graph of human tau levels (ELISA) in SEC-separated samples ( f ) Mean human tau levels of HMW (Frc. 2-4) and LMW (Frc. 13-16) SEC fractions. ( n =3-6) Unpaired t -test. ( g ) Immunoblot analysis (SDS-PAGE) of SEC-separated fractions from brain extracts (total tau, DAKO). Quantification of band density is also shown (right graphs) ( n =4). Unpaired t -test. ( h ) Dot blot analysis of PBS-soluble brain extracts with tau oligomer-specific antibody (T22), h

Supportive validation

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Experimental details: FIGURE 5 Investigation of early-life exposure to caffeine on hippocampal Tau phosphorylation and associated neuroinflammatory markers. (A) Western blot analysis of tau phosphorylation in water or caffeine exposed THY-Tau22 offsprings using antibodies targeting physiological (pThr181, Tau-1, pSer262, pSer396, pSer404) and pathologic (pThr212/Ser214) Tau epitopes. Quantifications were performed over total tau levels (Cter). Total tau levels were quantified versus GAPDH, used as loading control ( n = 7/group). (B) qPCR analysis of hippocampal neuroinflammatory markers associated with Tau pathology in the THY-Tau22 model ( n = 6-10 per group).