MN1000B

antibody from Invitrogen Antibodies

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

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Antibody data

Antibody Data
Antigen structure
References [93]
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Validations
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Product number: MN1000B - Provider product page
Provider: Invitrogen Antibodies
Product name: Tau Monoclonal Antibody (HT7), Biotin
Antibody type: Monoclonal
Antigen: Purifed from natural sources
Description: MN1000B targets Tau in ELISA, WB and IHC applications and shows reactivity with Bovine, and Human samples.
Conjugate: Biotin
Antibody clone number: HT7
Concentration: 0.1 mg/mL

MAPT protein structure - MN1000B shown in red.

Supportive validation

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

Supportive validation

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

Supportive validation

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

Supportive validation

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

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 4 Disulfide bond-mediated complex formation between tau and apoE in vitro. The in vitro interaction between tau and apoE proteins was evaluated by solution binding assay, followed by western blotting (four independent experiments). Recombinant human apoE protein was incubated with tau protein for 1 h at 37 degC in 20 ul of phosphate-buffered saline. The reaction was quenched by addition of 20 ul of sample buffer without (lane 1-7) or with (lane 8-10) reducing agent 2-mercaptoethanol (2-ME). a The presence of tau/apoE complexes (bands A and B) were determined by immunoblotting with antibodies to tau and apoE, respectively. The amounts of tau ( b ) and apoE ( c ) in the tau/apoE complexes were quantified. Bands A and B were tau/apoE complexes. Band C was a nonspecific band from tau immunoblot. Bands D and F were monomeric tau and apoE, respectively. Band E was estimated to be apoE dimer. Data are expressed as mean +- SEM relative to apoE2 condition. One-way ANOVA with Tukey post hoc tests were used. * P < 0.05; N.S. not significant
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 1 Tau protein in human vasculature. a Blood vessel isolations containing capillaries were validated by immunofluorescent labeling. DAPI-positive nuclei were also positive for endothelial cell markers such as von Willebrand factor (vWF) and zona occludins (ZO-1). b Isolates also contained basement membrane (collagen IV, ColIV) and some vessels were arteriolar in origin, indicated by the presence of smooth muscle actin (SMA). c Glut1 labeled endothelial cells were also occasionally observed to be surrounded by tau (HT7 antibody). d Western blotting of blood vessel isolations from human temporal cortex samples with varying degrees of tau pathology (Braak stage) indicates that vessels are frequently positive for tau (DAKO), but not other neuronal components (NeuN). Vessels are also enriched in Glut1 compared to total brain extracts. e When applied to a tau biosensor cell assay, vessels appear to retain their bioactivity with AD vessels exhibiting elevated seeding potential compared to controls (Mann-Whitney U test, p = 0.007, A.U. = arbitrary units). f Total frontal cortex protein from subjects with frontotemporal lobar dementia (FTLD) with tau exhibited greater bioactivity than control brains (Student's t test, p = 0.038). g Total tau protein in total brain versus blood vessel protein preparations as measured by ELISA. All graphs are plotted with means +/- standard deviations. * indicates p < 0.05, ** p < 0.01
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 2 Tau protein in vasculature from transgenic mice. a Tissue sections from rTg4510 or wild-type controls at 15 months of age were labeled for tau (HT7) and endothelial cells (Glut1). b An enlarged view of the box from ( a ) shows the extent of tau pathology at this age in transgenic (TG) and not wild-type (WT) mice. c Numerous blood vessels were observed to be closely associated with tau. d Western blotting of total cortex and isolated vessels indicates tau is present in vessels, but not other neuronal components such as NeuN. Glut1 is highly enriched. Vessels and cortex protein were run on the same blot, but separated here for clarity. e Tau bioactivity in a biosensor cell assay shows increased tau bioactivity in transgenic brain versus wild-type (Repeated Measures ANOVA, Sidak's post hoc, p = 0.003) and in transgenic blood vessels versus wild-type (Sidak's post hoc, p = 0.01). f Tau protein ELISA measures in total brain versus blood vessel protein. g Expression of the human Mapt transgene was observed by in situ hybridization in neurons but did not co-localize with Glut1-positive endothelial cells. Image is of a single plane captured using a confocal microscope. All graphs are plotted with means +/- standard deviations. * indicates p < 0.05, ** p < 0.01
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 3 Doxycycline treatment reduces soluble tau but does not alter vascular tau. a Sections from rTg4510 mice (TG) with and without doxycycline treatment were labeled for tau protein (HT7) showing a significant reduction in tau outside of neurofibrillary tangles. Insets show higher magnification images of cortical pathology. Scale bar = 10 mum. b Western blotting of human tau (HT7) present in Triton X (TX), sarkosyl soluble (SS) and sarkosyl insoluble (SI) fractions from a protein insolubility assay. c Quantification of western blots confirms reduced soluble tau present in TX (Student's t test, p = 0.001) and ( d ) SS fractions ( p = 0.0002) and ( e ) no change in SI fractions ( p = 0.31). Tau measures were normalized to a total protein stain to control for loading differences, which can be found along with uncropped blots in the supplement. f Western blotting of human tau in brain and isolated brain vessels from doxycycline treated (+) and untreated (-) mice. Glut1 is included to show enrichment of endothelial protein in vessels preparations. g Quantification of total human tau in vessels normalized to Glut1. h A biosensor cell assay shows retained tau bioactivity in transgenic blood vessels from Dox + mice (Repeated Measures ANOVA, Sidak's post hoc p = 0.04) and no difference in Dox- mice ( p = 0.89). All graphs are plotted with means +/- standard deviations. * indicates p < 0.05, *** p < 0.001
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 1 Tau and Abeta pathologies in the 5xFAD/AAV-tau mouse model. a Representative image of the distribution of Abeta (4G8, in green) and human tau (HT7, in red) in the hippocampal formation of 5xFAD/AAV-tau mice. b , c Higher magnification image in the ipsilateral ( b ) and contralateral CA3 ( c ). d Percentage of the total hippocampal area covered by HT7 staining in the ipsi- and contralateral hippocampus, in WT/AAV-tau and 5xFAD/AAV-tau mice. e Representative image of the distribution of Abeta (4G8, in green) and Ser202/Thr205 phospho-tau immunoreactivity (AT8, in red) in the hippocampus of 5xFAD/AAV-tau mice. f , g Higher magnification image in the ipsilateral ( f ) and contralateral CA3 ( g ). Note that in the ipsilateral hippocampus, AT8-positive tau is mainly located in the soma and neurites of neuronal cells. In the contralateral hippocampus, AT8-positive tau puncta (arrowheads) are mainly located near amyloid plaques (4G8, in green). h Percentage of the hippocampal surface covered by AT8 phospho-tau immunoreactivity in the ipsilateral hippocampus, comparing WT/AAV-tau and 5xFAD/AAV-tau mice. i Percentage of the surface covered by AT8 phospho-tau immunoreactivity in the contralateral hippocampus. Note the significant coverage increase in the 5xFAD/AAV-tau mice, which reflects the presence of phospho-tau-positive neuritic puncta. Scale bars: 800 um ( a , e ) and 100 um ( b , c , f , g ). Statistical analysis: unpaired two-tailed Student's t-test, **** p < 0.0001; WT/A
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 6 Effect of passive anti-Abeta immunization on human WT tau-induced pathological manifestations. a Representative images of human WT tau protein distribution (HT7, in red) in the ipsilateral side of the hippocampus, in a non-treated and a mAb11-treated 5xFAD/AAV-tau mouse. DAPI is shown in grey. Scale bars: 500 um. b Percentage of the ipsi- and contralateral hippocampal region covered by the HT7 staining, showing the effects of the mAb11-treatment in either 5xFAD/AAV-tau or WT/AAV-tau mice. Note the significant decrease of HT7 immunoreactivity in the ipsilateral hippocampus of 5xFAD/AAV-tau mice. c Representative images of the HT7-positive dystrophic neurites (red puncta indicated by arrowheads) near Abeta deposits (4G8, in green) in the contralateral hippocampus of a control sham-treated and a mAb11-treated 5xFAD/AAV-tau mouse. DAPI is shown in blue. Scale bars: 25 um. d Percentage of the Abeta-positive area (including a 5-um wide rim as described in Fig. 4 ) covered by the HT7 signal, which typically labels dystrophic neurites in the contralateral hippocampus of 5xFAD/AAV-tau mice. Note the significant decrease of the surface covered by human tau positive dystrophic neurites in the mAb11-treated mice. e Percentage of the ipsilateral hippocampus covered with AT8 positive signal in control sham-treated 5xFAD/AAV-tau mice compared to mAb11-treated mice. f Percentage of the plaque area (5-um extended Abeta plaque area as described in Fig. 4 ) covered by AT8 positive puncta
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 1 Syntenin-knock out reduces the AAV-mediated expression of tau-P301L in mouse brain. Micrographs of brain sections illustrating the expression of human tau protein after intracerebral injection of AAV-6-tau4R-P301L viral particles in the left hemisphere of wild-type mice (WT, top) or syntenin-knock out mice (KO, bottom). Note the low expression of tau in KO brain, both at caudal (-) and rostral (+) positions relative to the site of injection. Histogram on the right indicates the number of 'dark' pixels (with density above an arbitrarily set threshold) per complete brain section (left + right hemispheres). Data were collected using 4 mice per group and 4 sections per mice. Values are expressed as mean +- SEM. Scale bar is 1 mm.
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 1 Generation of Tau-KO and Tau-KD SH-SY5Y cells . a Scheme of the procedure used to generate CRISPR-Cas9-targeted cells and their characterization. Immune staining was performed with Tau13 antibody and nuclear staining with DAPI, western blot with Tau13 (loading control GAPDH) and immune precipitation and western blot with HT7 antibody, parental cells (wt) served as control. Amino acid sequences of the first MAPT coding exon in all lines demonstrate successful CRISPR-Cas9-editing causing frameshift (underlined in italics) into early stop codons (asterisks) for both alleles of 232P and 231K cells. b Scheme of procedure used to generate Tau-KD cell lines and their characterization by immune staining and western blot for Tau expression when compared to parental cells (wt) or cells transfected with the parental shRNA plasmid (ctrl). Scale bar 50 um.
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 2 PD-1 treatment did not enhance tau clearance. (A-F) PD-1 antibody treatment had no effect on soluble phospho-tau (PHF-1), total tau (Tau-5), total human tau (CP27) or ratios thereof on Western blots of tauopathy mouse brain homogenates, compared to control group. (A,G) PD-1 treatment had no effect on insoluble Western blot tau levels in the brain. (H,I) Enzyme-linked immunosorbent assay (ELISA) analyses of the same brain homogenates analyzed by Western blots confirmed lack of efficacy of the PD-1 therapy in clearing tau from the brain. Each bar represents the group average +/- SEM. As shown in individual scatterplots, 10 PD-1 treated- and eight control IgG mice were analyzed in each blot or ELISA assay. The total insoluble tau ELISA data failed normality test (IgG group) and was, therefore, analyzed by the Mann-Whitney test. All the other data passed the normality test and were, therefore, analyzed by unpaired t -test.
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 2 Expression of tau-degrading intrabodies decreases human tau in neurons. a Representative images of primary neurons co-expressing P301S-htau and the various anti-tau intrabodies. Expression of the conventional anti-tau intrabody, intrabody -K48R or the intrabody -K63R displayed a marked decrease in h-tau protein levels relative to the AAV-control; scale bar 50 mum. b P301S-htau protein quantification by tau ELISA in primary neurons co-expressing the various intrabodies revealed a significant decrease in h-tau protein levels upon expression of each tau-degrading intrabody relative to the AAV-controls. Expression of the conventional anti-tau intrabody displayed an non-significant decrease in h-tau protein relative to the AAV-control. Results are representative of three independent experiments, all data are expressed as mean +- s.e.m. and one-way ANOVA was performed with Tukey's Multiple Comparison. * p < 0.05
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 3 Immunofluorescence and fluorescence in situ analysis of anti-tau intrabody expression in aged P301S-tg mice. a Representative images of anti-HA for protein expression of the various anti-tau intrabodies in 9.5-month-old P301S-tg mice. Immunofluorescent detection of HA reveals clear-cut evidence of the non-targeting anti-tau intrabody within the ipsilateral hippocampus, whereas there was little to no immunofluorescence signal with the modified anti-tau intrabodies. Arrowhead indicating the injection site within the dentate gyrus (dg). b Representative images of in situ hybridization in 9.5-month-old P301S-tg mice validated the expression of the mRNA of the various anti-tau intrabodies within the hippocampus ipsilateral to the AAV injection. Scale bar 200 muM
Conjugate: Biotin