Anti-ER81/ETV1 antibody [DZR-1-29]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ER81/ETV1 antibody [DZR-1-29] (AB322139)

Immunohistochemical analysis of paraffin-embedded (A) U-87 MG (human glioblastoma-astrocytoma epithelial cell). (B) PC-3 (human prostate adenocarcinoma epithelial cell). (C) MCF7 (human breast adenocarcinoma epithelial cell). (D) T-47D (human breast adenocarcinoma epithelial cell). tissue labeling ER81/ETV1 with ab322139 at 1/500 (0.994 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining in (A) U-87 MG and (B) PC-3; no staining in (C) MCF7 and (D) T-47D.
The section was incubated with ab322139 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ER81/ETV1 antibody [DZR-1-29] (AB322139)

Immunohistochemical analysis of paraffin-embedded Human cerebral cortex tissue labeling ER81/ETV1 with ab322139 at 1/500 (0.994 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining in human cerebral cortex.
The section was incubated with ab322139 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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-ER81/ETV1 antibody [DZR-1-29] (AB322139)

Immunohistochemical analysis of paraffin-embedded Human cerebellum tissue labeling ER81/ETV1 with ab322139 at 1/500 (0.994 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining in human cerebellum.
The section was incubated with ab322139 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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-ER81/ETV1 antibody [DZR-1-29] (AB322139)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling ER81/ETV1 with ab322139 at 1/500 (0.994 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Low expression tissue : Nearly no staining on human colon.
The section was incubated with ab322139 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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-ER81/ETV1 antibody [DZR-1-29] (AB322139)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human embryonic kidney epithelial cell) transfected with a human ETV1 expression vector containing a His-tag. (B) HEK-293T transfected with a human ETV4 expression vector containing a His, Myc-tag. (C) HEK-293T transfected with a human ETV5 expression vector containing a His, Myc-tag. (D) HEK-293T transfected with an empty vector containing a His-tag. tissue labeling ER81/ETV1 with ab322139 at 1/10000 (0.05 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Nuclear staining in (A) HEK-293T transfected with a human ETV1 expression vector containing a His-tag; weak staining in sporadic cells of (B) HEK-293T transfected with a human ETV4 expression vector containing a His, Myc-tag and (C) HEK-293T transfected with a human ETV5 expression vector containing a His, Myc-tag. No staining in (D) HEK-293T transfected with an empty vector containing a His-tag.
The section was incubated with ab322139 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-ER81/ETV1 antibody [DZR-1-29] (AB322139)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat colon (perfused fixed) tissue labeling ER81/ETV1 with ab322139 at 1/50 (9.94 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).

Low expression : confocal image showing positive staining on periperal neurons of rat colon. The nuclear counterstain was DAPI (Blue). The section was incubated with ab322139 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

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

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ER81/ETV1 antibody [DZR-1-29] (AB322139)

Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling ER81/ETV1 with ab322139 at 1/500 (0.994 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : Nearly no staining on rat colon, only showed positive staining in sporadic peripheral neurons.
The section was incubated with ab322139 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

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-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-ER81/ETV1 antibody [DZR-1-29] (AB322139)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat spleen (perfused fixed) tissue labeling ER81/ETV1 with ab322139 at 1/50 (9.94 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).

Negative control : confocal image showing no staining on rat spleen. The nuclear counterstain was DAPI (Blue). The section was incubated with ab322139 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

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

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

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-ER81/ETV1 antibody [DZR-1-29] (AB322139)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat cerebellum (perfused fixed) tissue labeling ER81/ETV1 with ab322139 at 1/50 (9.94 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).

Panel A : merged staining of anti-ETV1 (ab322139, green), anti-NeuN (ab190565, grey) and anti-GFAP (ab201732, magenta) on rat cerebellum.
Panel B : anti-ETV1 stained on rat cerebellum.
Panel C : anti-NeuN stained in neurons of rat cerebellum.
Panel D : anti-GFAP stained in astrocytes of rat cerebellum.
The section was incubated in two rounds of staining : in the order of ab322139 and ab190565, ab201732 for 1 hr at room temperature. The nuclear counterstain was DAPI (Blue). The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

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

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ER81/ETV1 antibody [DZR-1-29] (AB322139)

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling ER81/ETV1 with ab322139 at 1/500 (0.994 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : Nearly no staining on mouse colon, only showed positive staining in sporadic peripheral neurons.
The section was incubated with ab322139 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

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-ER81/ETV1 antibody [DZR-1-29] (AB322139)

Immunohistochemical analysis of paraffin-embedded Mouse cerebellum tissue labeling ER81/ETV1 with ab322139 at 1/500 (0.994 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in mouse cerebellum.
The section was incubated with ab322139 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

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-ER81/ETV1 antibody [DZR-1-29] (AB322139)

Immunohistochemical analysis of paraffin-embedded Rat cerebellum tissue labeling ER81/ETV1 with ab322139 at 1/500 (0.994 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in rat cerebellum.
The section was incubated with ab322139 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

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-ER81/ETV1 antibody [DZR-1-29] (AB322139)

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

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

All lanes:

Western blot - Anti-ER81/ETV1 antibody [DZR-1-29] (ab322139) at 1/1000 dilution

Lane 1:

C6 (rat glial tumor glial cell) whole lysate at 20 µg

Lane 2:

Neuro-2a (mouse neuroblastoma neuroblast) whole lysate at 20 µg

Lane 3:

SH-SY5Y (human neuroblastoma epithelial cell) whole 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: 33-55 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-ER81/ETV1 antibody [DZR-1-29] (AB322139)

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

Low expression : MCF7, T-47D

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 18794142).

This blot was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

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

All lanes:

Western blot - Anti-ER81/ETV1 antibody [DZR-1-29] (ab322139) at 1/1000 dilution

Lane 1:

U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

T-47D (human ductal breast epithelial tumor epithelial cell) 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: 33-55 kDa,36 kDa

true

Exposure time: 125s