Anti-Tau 抗体 [EP2456Y]

• WB

Unknown

Western blot - Anti-Tau antibody [EP2456Y] (AB76128)

All lanes:

Western blot - Anti-Tau antibody [EP2456Y] (ab76128) at 1/10000 dilution

Lane 1:

Mouse brain lysate at 15 µg

Lane 2:

Rat brain lysate at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 78 kDa

false

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tau antibody [EP2456Y] (AB76128)

Immunohistochemical analysis of paraffin-embedded human liver labeling Tau with ab76128 at 1/2000 dilution, followed by a ready to use LeicaDS9800 (Bond^™ Polymer Refine Detection).

Negative Control : no staining on human liver.
The section was incubated with ab76128 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond^™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Tau antibody [EP2456Y] (AB76128)

Intracellular Flow Cytometry analysis of SH-SY5Y (Human neuroblastoma epithelial cell) cells labelling Tau with purified ab76128 at 1/500 dilution (1 μg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tau antibody [EP2456Y] (AB76128)

Immunohistochemical analysis of paraffin-embedded human cerebrum labeling Tau with ab76128 at 1/2000 dilution, followed by a ready to use LeicaDS9800 (Bond^™ Polymer Refine Detection).

Positive staining on human cerebrum.
The section was incubated with ab76128 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond^™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tau antibody [EP2456Y] (AB76128)

Immunohistochemical analysis of paraffin-embedded human astrocytoma labeling Tau with ab76128 at 1/2000 dilution, followed by a ready to use LeicaDS9800 (Bond^™ Polymer Refine Detection).

Positive staining on human astrocytoma.
The section was incubated with ab76128 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond^™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Tau antibody [EP2456Y] (AB76128)

Immunofluorescence staining of Tau using ab76128 (Unpurified format) in ioGlutamatergic Neurons (Human iPSC-Derived Glutamatergic Neurons, ab259259), which were differentiated for 11 days post induction.

The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% PBS-Tween for 5 mins and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab76128 at 0.5 μg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin, at 1/1000 dilution. Cells were then incubated with ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150120, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

Images were acquired with the Perkin Elmer Operetta HCA and a maximum intensity projection of confocal sections is shown.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tau antibody [EP2456Y] (AB76128)

Immunohistochemical analysis of paraffin-embedded mouse cerebrum labeling Tau with ab76128 at 1/2000 dilution, followed by a ready to use LeicaDS9800 (Bond^™ Polymer Refine Detection).

Positive staining on mouse cerebrum.
The section was incubated with ab76128 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond^™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tau antibody [EP2456Y] (AB76128)

Immunohistochemical analysis of paraffin-embedded rat cerebrum labeling Tau with ab76128 at 1/2000 dilution, followed by a ready to use LeicaDS9800 (Bond^™ Polymer Refine Detection).

Positive staining on rat cerebrum.
The section was incubated with ab76128 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond^™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Tau antibody [EP2456Y] (AB76128)

Immunocytochemistry analysis of Neuro-2a (Mouse neuroblastoma neuroblast) cells labeling Tau with purified ab76128 at 1/50 dilution (10 μg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% TritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1/200 (2.5 μg/mL). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/1000 (2 μg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• WB

Unknown

Western blot - Anti-Tau antibody [EP2456Y] (AB76128)

All lanes:

Western blot - Anti-Tau antibody [EP2456Y] (ab76128) at 1/20000 dilution

All lanes:

Human brain lysate at 15 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 78 kDa

false

• WB

Lab

Western blot - Anti-Tau antibody [EP2456Y] (AB76128)

Blocking/dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-Tau antibody [EP2456Y] (ab76128) at 1/1000 dilution

Lane 1:

293T cells transfected with an empty vector containing a flag tag whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a human Tau expression vector containing a flag whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a human MAP2 expression vector containing a flag whole cell lysate at 20 µg

Lane 4:

293T cells transfected with a human MAP4 expression vector containing a flag whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 78 kDa

Observed band size: 55-100 kDa

false

Exposure time: 1s

• WB

Lab

Western blot - Anti-Tau antibody [EP2456Y] (AB76128)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

In Western blot, Anti-Tau antibody [EP2456Y] (ab76128) Total Protein Control at 1/1000 dilution.

All lanes:

Western blot - Anti-Tau (phospho S324) antibody [EPR2457(2)] (<a href='/products/primary-antibodies/tau-phospho-s324-antibody-epr24572-ab109401'>ab109401</a>) at 1/1000 dilution

Lane 1:

Untreated mouse brain tissue lysate (untreated membrane) at 20 µg

Lane 2:

Mouse brain treated with 1μM Okadaic Acid and 200nM Calyculin A for 1 hour, tissue lysate (untreated membrane) at 20 µg

Lane 3:

Untreated mouse brain tissue lysate (alkaline phosphatase treated membrane) at 20 µg

Lane 4:

Mouse brain treated with 1μM Okadaic Acid and 200nM Calyculin A for 1 hour, tissue lysate (alkaline phosphatase treated membrane) at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 48-60 kDa

false

Exposure time: 136s

• WB

Lab

Western blot - Anti-Tau antibody [EP2456Y] (AB76128)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

In Western blot, Anti-Tau antibody [EP2456Y] (ab76128) Total Protein Control at 1/1000 dilution.

All lanes:

Western blot - Anti-Tau (phospho S324) antibody [EPR2457(2)] (<a href='/products/primary-antibodies/tau-phospho-s324-antibody-epr24572-ab109401'>ab109401</a>) at 1/1000 dilution

Lane 1:

Untreated rat brain tissue lysate (untreated membrane) at 20 µg

Lane 2:

Rat brain treated with 1μM Okadaic Acid and 200nM Calyculin A for 1 hour, tissue lysate (untreated membrane) at 20 µg

Lane 3:

Untreated rat brain tissue lysate (alkaline phosphatase treated membrane) at 20 µg

Lane 4:

Rat brain treated with 1μM Okadaic Acid and 200nM Calyculin A for 1 hour, tissue lysate (alkaline phosphatase treated membrane) at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 48-60 kDa

false

Exposure time: 70s

• WB

Lab

Western blot - Anti-Tau antibody [EP2456Y] (AB76128)

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Anti-Tau antibody [EP2456Y] (ab76128) - Total Protein Control (ab76128) at 1/1000 dilution.

All lanes:

Western blot - Anti-Tau (phospho S235) antibody [EPR2452] (<a href='/products/primary-antibodies/tau-phospho-s235-antibody-epr2452-ab133253'>ab133253</a>) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate (untreated membrane) at 20 µg

Lane 2:

Rat brain tissue lysate (untreated membrane) at 20 µg

Lane 3:

Mouse brain tissue lysate (alkaline phosphatase treated membrane) at 20 µg

Lane 4:

Rat brain tissue lysate (alkaline phosphatase treated membrane) at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 35-80 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Tau antibody [EP2456Y] (AB76128)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 22366723, PMID : 24113653).

The expression of Tau (O-GlcNAc S400) is upregulated in response to Thiamet G treatment (PMID : 25336656, PMID : 22366723, PMID : 24113653).

The identity of the higher MW band at approximately 110 kDa is unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

In Western blot, Anti-Tau antibody - Total protein control (ab76128) staining at 1/1000 dilution.

In Western blot, Anti-Tau antibody - Total protein control Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution

All lanes:

Western blot - Anti-Tau (O-GlcNAc S400) antibody [EPR24891-48] (<a href='/products/primary-antibodies/tau-o-glcnac-s400-antibody-epr24891-48-ab320076'>ab320076</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with a human wild-type Tau 2N4R expression vector containing a myc-His-tag®, whole cell lysate at 50 µg

Lane 2:

293T cells transfected with a human wild-type Tau 2N4R expression vector containing a myc-His-tag® were treated with 10 uM Thiamet G for 48 hours, whole cell lysate at 50 µg

Lane 3:

293T cells transfected with a human Tau 2N4R (S400A mutation) expression vector containing a myc-His-tag®, whole cell lysate at 50 µg

Lane 4:

293T cells transfected with a human Tau 2N4R (S400A mutation) expression vector containing a myc-His-tag® were treated with 10 uM Thiamet G for 48 hours, whole cell lysate at 50 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 25-75 kDa,36 kDa

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Anti-Tau antibody [EP2456Y] (AB76128)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

In Western blot, Anti-Tau antibody [EP2456Y] - (ab76128) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-Tau (acetyl K311) antibody [EPR29545-504] (<a href='/products/primary-antibodies/tau-acetyl-k311-antibody-epr29545-504-ab324728'>ab324728</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a myc-His-tag, whole cell lysate at 4 µg

Lane 2:

293T cells transfected with a human 2N4R Tau (WT) expression vector containing a myc-His-tag®, whole cell lysate at 4 µg

Lane 3:

293T cells co-transfected with a human 2N4R Tau (WT) expression vector containing a myc-His-tag® and a human CBP HAT domain expression vector containing a myc-His-tag® whole cell lysate at 4 µg

Lane 4:

293T cells transfected with a human 2N4R Tau (K311R) expression vector containing a myc-His-tag®, whole cell lysate at 4 µg

Lane 5:

293T cells co-transfected with a human 2N4R Tau (K311R) expression vector containing a myc-His-tag® and a human CBP HAT domain expression vector containing a myc-His-tag®, whole cell lysate at 4 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 70 kDa,36 kDa

false

Exposure time: 180s

• WB

CiteAb

Western blot - Anti-Tau antibody [EP2456Y] (AB76128)

Tau western blot using anti-Tau antibody [EP2456Y] ab76128. Publication image and figure legend from Chakravarthy, M., Chen, S., et al., 2020, Genes (Basel), PubMed 32575375.

ab76128 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab76128 please see the product overview.

(A) Densitometry analysis of the tau expression obtained from western blot (three replicates) analysis after the treatment of SH-SY5Y cells with AO4 at 50 nM concentrations and incubation of AO4 for 24 h, 48 h, and six days. SH-SY5Y cells were treated with the scrambled control at a 50 nM concentration and incubation of six days. AO4 targets exon 4 of the MAPT transcript. The error bars represent the standard error of the mean. (B) Densitometry analysis of the tau expression obtained from western blot (three replicates) analysis after the treatment of SH-SY5Y cells with RNV563 at 400 nM concentrations and incubation of DNAzyme for 24 h, 48 h, and six days. SH-SY5Y cells were treated with the scrambled control at a 400 nM concentration and incubation of six days. RNV563 targets exon 9 of the MAPT transcript. The error bars represent the standard error of the mean.

false