Anti-1N Tau antibody [EPR25206-82]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-1N Tau antibody [EPR25206-82] (AB314889)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling 1N Tau with ab314889 at 1/2000 (0.255 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human cerebrum.The section was incubated with ab314889 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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-1N Tau antibody [EPR25206-82] (AB314889)

Immunohistochemical analysis of paraffin-embedded Human hippocampus tissue labeling 1N Tau with ab314889 at 1/2000 (0.255 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human hippocampus.The section was incubated with ab314889 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

• WB

Supplier Data

Western blot - Anti-1N Tau antibody [EPR25206-82] (AB314889)

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

Negative control : spleen (PMID : 24386422).

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

All lanes:

Western blot - Anti-1N Tau antibody [EPR25206-82] (ab314889) at 1/1000 dilution

Lane 1:

Human hippocampus tissue lysate at 80 µg

Lane 2:

Human spleen tissue lysate at 80 µg

Secondary

All lanes:

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

Observed band size: 55 kDa,36 kDa

true

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-1N Tau antibody [EPR25206-82] (AB314889)

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

Negative control : spleen (PMID : 24386422).

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

All lanes:

Western blot - Anti-1N Tau antibody [EPR25206-82] (ab314889) at 1/1000 dilution

Lane 1:

Rat hippocampus tissue lysate at 80 µg

Lane 2:

Rat spleen tissue lysate at 80 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/10000 dilution

Observed band size: 55 kDa,15 kDa

false

Exposure time: 59s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-1N Tau antibody [EPR25206-82] (AB314889)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human embryonic kidney epithelial cell) transfected with a human Tau 1N3R expression vector containing a his tag. (B) HEK-293T transfected with a human Tau 1N4R expression vector containing a his tag. (C) HEK-293T transfected with a human Tau 0N3R expression vector containing a his tag. (D) HEK-293T transfected with a human Tau 2N3R expression vector containing a his tag. (E) HEK-293T transfected with a human Tau 0N4R expression vector containing a his tag. (F) HEK-293T transfected with a human Tau 2N4R expression vector containing a his tag. (G) HEK-293T cells transfected with empty vector containing a his tag. tissue labeling 1N Tau with ab314889 at 1/20000 (0.026 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) HEK-293T transfected with a human Tau 1N3R expression vector containing a his tag and (B) HEK-293T transfected with a human Tau 1N4R expression vector containing a his tag. No staining on (C) HEK-293T transfected with a human Tau 0N3R expression vector containing a his tag, (D) HEK-293T transfected with a human Tau 2N3R expression vector containing a his tag, (E) HEK-293T transfected with a human Tau 0N4R expression vector containing a his tag, (F) HEK-293T transfected with a human Tau 2N4R expression vector containing a his tag, and (G) HEK-293T cells transfected with empty vector containing a his tag. The section was incubated with ab314889 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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-1N Tau antibody [EPR25206-82] (AB314889)

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling 1N Tau with ab314889 at 1/2000 (0.255 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on human spleen. The section was incubated with ab314889 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-1N Tau antibody [EPR25206-82] (AB314889)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) cells labelling 1N Tau with ab314889 at 1/50 (10.2 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing positive staining in 293T cells transfected with a human Tau 1N3R and Tau 1N4R expression vector containing a myc-tag, showing no staining in Tau 0N4R/0N3R/2N3R/2N4R(WT) and empty expression vector containing a myc-tag. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

ab223894 Anti-Myc tag mouse monoclonal antibody (Alexa Fluor® 594) was used to counterstain at 1/100 (5ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• WB

Supplier Data

Western blot - Anti-1N Tau antibody [EPR25206-82] (AB314889)

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] (ab314889) 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='/en-us/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-1N Tau antibody [EPR25206-82] (AB314889)

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] (ab314889) 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='/en-us/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