44-764G

antibody from Invitrogen Antibodies

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

Provider product page for 44-764G

Western blot

Other assay

Antibody data

Antibody Data
Antigen structure
References [23]
Comments [0]
Validations
Western blot [2]
Other assay [17]

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Product number: 44-764G - Provider product page
Provider: Invitrogen Antibodies
Product name: Phospho-Tau (Ser422) Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Synthetic peptide
Reactivity: Human
Host: Rabbit
Isotype: IgG
Vial size: 100 µL
Storage: -20°C

MAPT protein structure - 44-764G shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Up-regulation and Antibody-Peptide Competition. Human recombinant Tau stimulated with GSK-3β (1 µg per µg Tau) for 45 minutes was added to background extracts, resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. The membrane was blocked with a 5% BSA-TBST buffer for one hour at room temperature and either left untreated (1-4) or treated with lambda phosphatase (5), then incubated with the Tau (pS422) antibody in a 3% BSA-TBST buffer for two hours at room temperature, following prior incubation with: no peptide (1, 5), the non-phosphopeptide corresponding to the phosphopeptide immunogen (2), a generic phosphoserine-containing peptide (3), or the phosphopeptide immunogen (4). After washing, the membrane was incubated with goat F (ab')2 anti-rabbit IgG HRP conjugate (Product # ALI4404) and signals were detected using the Pierce SuperSignal™ method.The data show that only the phosphopeptide corresponding to Tau (pS422) blocks the antibody signal, demonstrating the specificity of the antibody. The data also show that phosphatase stripping eliminates the signal, verifying that the antibody is phospho-specific. (Product # 44-764G)

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Up-regulation and Antibody-Peptide Competition. Human recombinant Tau stimulated with GSK-3β (1 µg per µg Tau) for 45 minutes was added to background extracts, resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. The membrane was blocked with a 5% BSA-TBST buffer for one hour at room temperature and either left untreated (1-4) or treated with lambda phosphatase (5), then incubated with the Tau (pS422) antibody in a 3% BSA-TBST buffer for two hours at room temperature, following prior incubation with: no peptide (1, 5), the non-phosphopeptide corresponding to the phosphopeptide immunogen (2), a generic phosphoserine-containing peptide (3), or the phosphopeptide immunogen (4). After washing, the membrane was incubated with goat F (ab')2 anti-rabbit IgG HRP conjugate (Product # ALI4404) and signals were detected using the Pierce SuperSignal™ method.The data show that only the phosphopeptide corresponding to Tau (pS422) blocks the antibody signal, demonstrating the specificity of the antibody. The data also show that phosphatase stripping eliminates the signal, verifying that the antibody is phospho-specific. (Product # 44-764G)

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: 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: 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: 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 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: 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

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 3 In vitro biochemical interaction between PTK2B and Tau. ( a ) Pull down of PTK2B with GST, GST-Tau1N4R and GST-Tau2N4R protein constructs. Upper panel: detection of PTK2B in the pull down extract by western blotting. Lower panel: corresponding Coomassie blue gel used as loading control of GST constructs. ( b ) Pull down of Tau1N4R and Tau2N4R with GST and GST-PTK2B protein constructs. Upper panel: detection of Tau (arrows) in the pull down extract by western blotting (an unspecific band is labeled with a star, MW, molecular weight). Lower panel: corresponding Coomassie blue gel used as loading control of GST constructs. As the GST-PTK2B constructs were difficult to produce and visualize with Coomassie blue staining (arrows), we checked the GST-PTK2B construct through detection of PTK2B by western blotting (middle panel). All results shown in panels ( a and b ) are representative of three independent experiments. ( c ) Western blotting analysis of Tau phosphorylation in different Fak conditions in the Drosophila eye. Actin is used as a loading control ( n =2-4). ( d ) Images and quantification of fly eyes co-expressing the 2N4R Tau isoform and a mCD8::GFP construct used as control or wild-type Fak or mutant Fak Y430F (scale bar 0.1 mm).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
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

Submitted by: Invitrogen Antibodies (provider)
Main image
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

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig 6. Brain sections of WT, APP and Tau mice stained by antibodies specific to A beta (6E10) (a), phosphorylated tau: Tau-pS422 (b), reactive astrocytes (GFAP) (c), and microglia (IBA1) (d). Scale bars are 200 mu m in (a-c) and 100 mu m in (d).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
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