44-738G

antibody from Invitrogen Antibodies

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

Provider product page for 44-738G

Western blot

Immunocytochemistry

Immunohistochemistry

Flow cytometry

Other assay

Antibody data

Antibody Data
Antigen structure
References [33]
Comments [0]
Validations
Western blot [1]
Immunocytochemistry [1]
Flow cytometry [1]
Other assay [16]

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Product number: 44-738G - Provider product page
Provider: Invitrogen Antibodies
Product name: Phospho-Tau (Thr205) Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Synthetic peptide
Description: Purified from rabbit serum by sequential epitope-specific chromatography, this product contains enough material for 10 mini-blots. This 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 threonine 205. The antibody has been used in western blotting. Previous lots of this antibody have been used in immunohistochemistry. In western analysis the positive control used was recombinant human Tau treated with GSK3beta.
Reactivity: Human, Mouse, Rat
Host: Rabbit
Isotype: IgG
Vial size: 100 µL
Storage: -20°C

MAPT protein structure - 44-738G shown in red.

Supportive validation

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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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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

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig 5 Tau phosphorylation is decreased in neurons treated with 20 mM KCl. Immunoblot analysis of Tau phosphorylation in neurons treated either with 10mM or 20mM KCl. The phosphorylation of S199/S202, T181, T205 and S404 was not significantly affected by 10mM of KCl treatment whereas the phosphorylation of all these sites was decreased by 20mM of KCl treatment. The signal of the phospho-antibodies was normalized to that of total Tau (n = 5, mean +- SEM, unpaired t-test two-tailed, * P < 0.05, *** P < 0.001). The amount of dephosphorylated Tau revealed by the Tau-1 antibody was normalized to that of total Tau lysate (n = 5, mean +- SEM, unpaired t-test two-tailed, * P < 0.05).

Supportive validation

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Experimental details: Figure 1 Tau is rapidly dephosphorylated site-specifically in mouse brains during postmortem. Mice were scarified by cervical dislocation. The dead animal bodies were kept at room temperature for the indicated periods of time. ( A ) Phosphorylation of tau was analyzed by Western blots developed with phosphorylation-dependent and site-specific tau antibodies indicated at the right side of the blots. ( B ) The levels of tau phosphorylation at individual sites were quantified and normalized with total tau level and presented as mean +- SEM. (n = 3-4). * p < 0.05; ** p < 0.01.

Supportive validation

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Experimental details: Figure 2 Both HMW-tau and LMW-tau are selectively hyperphosphorylated in AD brain. (A) AD and control human brain homogenates were analyzed by western blots developed with the indicated site-specific and phosphorylation dependent anti-tau antibodies. (B,C) Blots were analyzed by densitometry. The levels of hyperphosphorylated HMW-tau (B) and LMW-tau (C) are shown as scattered dots with mean +- SD. ** p < 0.01; *** p < 0.001; **** p < 0.0001.

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: Fig. 3 Imaging of CDK5 substrate phosphorylaiton in primary cells. Rat primary cortical neurons were cultured for 6 days in vitro. a Cells were incubated with 10 muM purvalanol A or vehicle for 3 h prior to fixation, permeabolisation and staining with the indicated antibodies. Phosphospecific antibodies were detected by Cy-3 bound 2ry antibodies ( red ) and nuclei were counter-stained with the DNA-binding dye DAPI ( blue ). Scale bar = 60 mum. b The primary neuronal cultures were incubated with either 10 muM purvalanol A, roscovitine or vehicle for 3 h prior to fixation in formaldehyde and embedding in paraffin. Sections were taken from each paraffin block and incubated with the phospho-antibodies as indicated, then biotinylated secondary antibody followed by streptavidin complexed with biotinylated peroxidases which were visualized using DAB staining. Cell nuclei were counterstained with hematoxylin and mounted in DPX. Scale bar = 100 mum. Images are representative of sets from at least three different neuron preparations

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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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