Anti-Tau 抗体 [EPR22524-95]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tau antibody [EPR22524-95] (AB254256)

Immunohistochemical analysis of paraffin-embedded human Alzheimer's brain tissue* labelling Tau with ab254256 at 0.1ug/ml followed by a ready to use LeicaDS9800 (Bond^™ Polymer Refine Detection).

Positive staining in human Alzheimer's brain.

The section was incubated with ab254256 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 with sodium citrate buffer (pH 6.0 epitope retrieval solution1) for 20 mins.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tau antibody [EPR22524-95] (AB254256)

Immunohistochemical analysis of paraffin-embedded human cerebral cortex tissue labeling Tau with ab254256 at 1/4000 dilution, followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human cerebral cortex (PMID : 22961084) is observed. Counterstained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

The section was incubated with ab254256 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• IP

Unknown

Immunoprecipitation - Anti-Tau antibody [EPR22524-95] (AB254256)

Tau was immunoprecipitated from 0.35 mg human brain lysate with ab254256 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab254256 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used at 1/5000 dilution.

Lane 1 : Human brain lysate 10 μg (Input).
Lane 2 : ab254256 IP in human brain lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab254256 in human brain lysate.

Blocking/Dilution buffer : 5% NFDM/TBST.
Exposure time : 10 seconds.

All lanes:

Immunoprecipitation - Anti-Tau antibody [EPR22524-95] (ab254256)

Predicted band size: 78 kDa

Observed band size: 50 kDa

false

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tau antibody [EPR22524-95] (AB254256)

Immunohistochemical analysis of paraffin-embedded mouse hippocampus tissue labeling Tau with ab254256 at 1/4000 dilution, followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse hippocampus (PMID : 22961084) is observed. Counterstained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

The section was incubated with ab254256 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-Tau antibody [EPR22524-95] (AB254256)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse cerebral cortex tissue labeling Tau with ab254256 at 1/100 dilution (green), followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at a 1/1,000 dilution. Positive staining on mouse cerebral cortex (PMID : 22961084) is observed. Counterstained with DAPI (blue).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ab150077 AlexaFluor^®488 Goat anti-Rabbit used at a 1/1,000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10 mM citrate pH 6.0 and 0.05% Tween-20).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tau antibody [EPR22524-95] (AB254256)

Immunohistochemical analysis of paraffin-embedded mouse cerebral cortex tissue labeling Tau with ab254256 at 1/4000 dilution, followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse cerebral cortex (PMID : 22961084) is observed. Counterstained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

The section was incubated with ab254256 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• WB

Lab

Western blot - Anti-Tau antibody [EPR22524-95] (AB254256)

Blocking/dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-Tau antibody [EPR22524-95] (ab254256) 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

Unknown

Western blot - Anti-Tau antibody [EPR22524-95] (AB254256)

Negative control : human fetal heart, human fetal kidney, human spleen. (PMID : 24386422).

The molecular weight observed is consistent with what has been described in the literature (PMID : 24386422).

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Tau antibody [EPR22524-95] (ab254256) at 1/1000 dilution

Lane 1:

Human brain lysate at 10 µg

Lane 2:

Human fetal heart lysate at 10 µg

Lane 3:

Human fetal kidney lysate at 10 µg

Lane 4:

Human spleen lysate at 10 µg

Secondary

All lanes:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Predicted band size: 78 kDa

Observed band size: 50-70 kDa

false

Exposure time: 8s

• WB

Supplier Data

Western blot - Anti-Tau antibody [EPR22524-95] (AB254256)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. Tau assembles into oligomers as described in PMID : 28382304, 32692785 and 30120733. In Western blot, anti-Tau antibody (ab254256) loading control staining at 1/1000 dilution. Exposure time : 26 seconds.

All lanes:

Western blot - Anti-Tau (phospho S396) antibody [E178] - Mouse IgG2a (Chimeric) - BSA and Azide free (<a href='/products/primary-antibodies/tau-phospho-s396-antibody-e178-mouse-igg2a-chimeric-bsa-and-azide-free-ab302491'>ab302491</a>) at 1/1000 dilution

Lane 1:

Rat brain tissue lysate (untreated membrane) at 10 µg

Lane 2:

Rat brain tissue lysate (phosphatase treated membrane) at 10 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/5000 dilution

Observed band size: 50-60 kDa,200 kDa

false

• WB

Supplier Data

Western blot - Anti-Tau antibody [EPR22524-95] (AB254256)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. In Western blot, anti-Tau antibody (ab254256) loading control staining at 1/1000 dilution. Exposure time : 3.25 seconds.

All lanes:

Western blot - Anti-Tau (phospho S396) antibody [E178] - Mouse IgG2a (Chimeric) - BSA and Azide free (<a href='/products/primary-antibodies/tau-phospho-s396-antibody-e178-mouse-igg2a-chimeric-bsa-and-azide-free-ab302491'>ab302491</a>) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate (untreated membrane) at 10 µg

Lane 2:

Mouse brain tissue lysate (phosphatase treated membrane) at 10 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/5000 dilution

Observed band size: 50-70 kDa

false

• WB

Unknown

Western blot - Anti-Tau antibody [EPR22524-95] (AB254256)

Negative control : mouse heart, mouse kidney, mouse spleen. (PMID : 24386422).

The molecular weight observed is consistent with what has been described in the literature (PMID : 24386422).

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Tau antibody [EPR22524-95] (ab254256) at 1/1000 dilution

Lane 1:

Mouse brain lysate at 10 µg

Lane 2:

Mouse heart lysate at 10 µg

Lane 3:

Mouse kidney lysate at 10 µg

Lane 4:

Mouse spleen lysate at 10 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

Predicted band size: 78 kDa

Observed band size: 50-70 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-Tau antibody [EPR22524-95] (AB254256)

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

In Western blot, Anti-Tau antibody [EPR22524-95] (ab254256) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-1N Tau antibody [EPR25206-82] (<a href='/products/primary-antibodies/1n-tau-antibody-epr25206-82-ab314889'>ab314889</a>) at 1/1000 dilution

Lane 1:

His-tagged Tau 0N3R human recombinant protein at 50 ng

Lane 2:

His-tagged Tau 0N4R human recombinant protein at 50 ng

Lane 3:

His-tagged Tau 1N3R human recombinant protein at 10 ng

Lane 4:

His-tagged Tau 1N4R human recombinant protein at 10 ng

Lane 5:

His-tagged Tau 2N3R human recombinant protein at 50 ng

Lane 6:

His-tagged Tau 2N4R human recombinant protein at 50 ng

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: 75 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Tau antibody [EPR22524-95] (AB254256)

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

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) staining at 1/100000 dilution.

In Western blot, Anti-6X His tag antibody [EPR20547]-CHIP Grade (ab213204) staining at 1/5000 dilution, Anti-Tau antibody [EPR22524-95] (ab254256) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-1N Tau antibody [EPR25206-82] (<a href='/products/primary-antibodies/1n-tau-antibody-epr25206-82-ab314889'>ab314889</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

293T cells transfected with a human Tau 0N3R expression vector containing a His-tag, whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a human Tau 0N4R expression vector containing a His-tag, whole cell lysate at 20 µg

Lane 4:

293T cells transfected with a human Tau 1N3R expression vector containing a His-tag, whole cell lysate at 20 µg

Lane 5:

293T cells transfected with a human Tau 1N4R expression vector containing a His-tag, whole cell lysate at 20 µg

Lane 6:

293T cells transfected with a human Tau 2N3R expression vector containing a His-tag, whole cell lysate at 20 µg

Lane 7:

293T cells transfected with a human Tau 2N4R expression vector containing a His-tag, 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 1/100000 dilution

Observed band size: 37-75 kDa,15 kDa

false

Exposure time: 6s

• WB

Supplier Data

Western blot - Anti-Tau antibody [EPR22524-95] (AB254256)

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

In Western blot, Anti-Tau antibody - Total protein control (ab254256) staining at 1/1000 dilution.

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

All lanes:

Western blot - Anti-Tau (nitrosyl Y29) antibody [EPR27841-79] (<a href='/products/primary-antibodies/tau-nitrosyl-y29-antibody-epr27841-79-ab315891'>ab315891</a>) at 1/1000 dilution

Lane 1:

Untreated 293T cells transfected with an empty vector containing a His-tag, whole cell lysate at 20 µg

Lane 2:

293T cells transfected with an empty vector containing a His-tag were treated with 3mM peroxynitrite for 1 hour, whole cell lysate at 20 µg

Lane 3:

Untreated 293T cells transfected with a human Tau 2N4R expression vector containing a His-tag, whole cell lysate at 20 µg

Lane 4:

293T cells transfected with a human Tau 2N4R expression vector containing a His-tag were treated with 3mM peroxynitrite for 1 hour, 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 1/100000 dilution

Observed band size: 20-75 kDa,36 kDa

false

Exposure time: 180s