Anti-Angiotensin II Type 2 Receptor antibody [EPR30232-537] - BSA and Azide free

• ICC

Supplier Data

Immunocytochemistry - Anti-Angiotensin II Type 2 Receptor antibody [EPR30232-537] - BSA and Azide free (AB326595)

This data was developed using ab324679, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized RD (human muscle multinucleated spindle-shaped cell) cells labelling Angiotensin II Type 2 Receptor with ab324679 at 1/50 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing membranous staining in RD cell line and no staining in HeLa cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Low expression : HeLa.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 (1 ug/ml) dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 (2 ug/ml) dilution (Magenta).

-ve control 1 : ab324679 at a 1/50 dilution followed by ab150120 at a 1/1000 dilution.

-ve control 2 : ab7291 at a 1/1000 dilution followed by ab150081 at a 1/1000 dilution.

• IHC

Supplier Data

Immunohistochemistry - Anti-Angiotensin II Type 2 Receptor antibody [EPR30232-537] - BSA and Azide free (AB326595)

This data was developed using ab324679, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat adrenal gland tissue labeling Angiotensin II Type 2 Receptor with ab324679 at 1/100 (5.0 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat adrenal medulla.
The section was incubated with ab324679 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 citrate buffer (pH 9.0 epitope retrieval solution 2) for 20 mins

• IHC

Supplier Data

Immunohistochemistry - Anti-Angiotensin II Type 2 Receptor antibody [EPR30232-537] - BSA and Azide free (AB326595)

This data was developed using ab324679, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse adrenal gland tissue labeling Angiotensin II Type 2 Receptor with ab324679 at 1/100 (5.0 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse adrenal medulla.
The section was incubated with ab324679 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 citrate buffer (pH 9.0 epitope retrieval solution 2) for 20 mins

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Angiotensin II Type 2 Receptor antibody [EPR30232-537] - BSA and Azide free (AB326595)

This data was developed using ab324679, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse adrenal gland tissue staining Angiotensin II Type 2 Receptor with ab324679 at a 1/100 ( 5.0 µg/ml) dilution ab308287 anti-VGF used at a 1/500 ( 0.498 µg/ml) dilution and ab324019 anti-CYP21A2 used at a 1/5000 ( 0.1 µg/ml) dilution.

Panel A : merged staining of anti-Angiotensin II Type 2 Receptor (green; Opal™520) anti-VGF (magenta; Opal™570) and anti-CYP21A2 (gray; Opal™690) on mouse adrenal gland.

Panel B : anti-Angiotensin II Type 2 Receptor staining membrane of adrenal medulla in mouse adrenal gland.

Panel C : anti-VGF staining endocrine cells of adrenal medulla in mouse adrenal gland.

Panel D : anti-CYP21A2 staining adrenal cortex in mouse adrenal gland.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab324679 ab308287 and ab324019 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

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

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Angiotensin II Type 2 Receptor antibody [EPR30232-537] - BSA and Azide free (AB326595)

This data was developed using ab324679, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded rat adrenal gland tissue staining Angiotensin II Type 2 Receptor with ab324679 at a 1/100 ( 5.0 µg/ml) dilution ab308287 anti-VGF used at a 1/500 ( 0.498 µg/ml) dilution and ab324019 anti-CYP21A2 used at a 1/5000 ( 0.1 µg/ml) dilution.

Panel A : merged staining of anti-Angiotensin II Type 2 Receptor (green; Opal™520) anti-VGF (magenta; Opal™570) and anti-CYP21A2 (gray; Opal™690) on rat adrenal gland.

Panel B : anti-Angiotensin II Type 2 Receptor staining membrane of adrenal medulla in rat adrenal gland.

Panel C : anti-VGF staining endocrine cells of adrenal medulla in rat adrenal gland.

Panel D : anti-CYP21A2 staining adrenal cortex in rat adrenal gland.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab324679 ab308287 and ab324019 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

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-Angiotensin II Type 2 Receptor antibody [EPR30232-537] - BSA and Azide free (AB326595)

This data was developed using ab324679, the same antibody clone in a different buffer formulation.

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

Low expression : liver skin

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 31511323).

Samples are non-boiled as boiling may cause protein aggregation.

Lanes 1-2 are incubated with Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 and lanes 3-6 are incubated with Goat Anti-Rabbit IgG (H+L) Peroxidase conjugated (ab97051) at 1/20000.

The identity of the higher MW band above 250 kDa in lane 2 is unknow.

The identity of the bands lower than 37 kDa in lanes 3-6 are unknown.

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

All lanes:

Western blot - Anti-Angiotensin II Type 2 Receptor antibody [EPR30232-537] (<a href='/en-us/products/primary-antibodies/angiotensin-ii-type-2-receptor-antibody-epr30232-537-ab324679'>ab324679</a>) at 1/1000 dilution

Lane 1:

Human uterus tissue lysate at 40 µg

Lane 2:

Human liver tissue lysate at 40 µg

Lane 3:

Rat adrenal gland tissue lysate at 40 µg

Lane 4:

Rat uterus tissue lysate at 40 µg

Lane 5:

Rat skin tissue lysate at 40 µg

Lane 6:

Rat liver tissue lysate at 40 µg

Secondary

Lanes 1 - 6:

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 1/2000 dilution

Lanes 1 - 6:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/20000 dilution

Observed band size: 50-100 kDa,36 kDa

false

Exposure time: 103s

• WB

Supplier Data

Western blot - Anti-Angiotensin II Type 2 Receptor antibody [EPR30232-537] - BSA and Azide free (AB326595)

This data was developed using ab324679, the same antibody clone in a different buffer formulation.

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

Low expression : HeLa SK-N-BE(2) HepG2

BFA prevents complex glycosylation of target a fraction of which can reach the cell surface. The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 31511323).

Samples are non-boiled as boiling may cause protein aggregation.

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

Exposure time : Lanes 1-4 : 59 seconds; Lanes 5-6 : 26 seconds

All lanes:

Western blot - Anti-Angiotensin II Type 2 Receptor antibody [EPR30232-537] (<a href='/en-us/products/primary-antibodies/angiotensin-ii-type-2-receptor-antibody-epr30232-537-ab324679'>ab324679</a>) at 1/1000 dilution

Lane 1:

RD (human muscle multinucleated spindle-shaped cell) whole cell lysate at 40 µg

Lane 2:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 40 µg

Lane 3:

SK-N-BE(2) (human neuroblastoma neuroblast) whole cell lysate at 40 µg

Lane 4:

HepG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate at 40 µg

Lane 5:

Untreated RD (human muscle multinucleated spindle-shaped cell) whole cell lysate at 40 µg

Lane 6:

RD treated with /ml BFA for 24 hours whole cell lysate at 40 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 1/20000 dilution

Observed band size: 50-100 kDa,36 kDa

false

• IHC

Supplier Data

Immunohistochemistry - Anti-Angiotensin II Type 2 Receptor antibody [EPR30232-537] - BSA and Azide free (AB326595)

This data was developed using ab324679, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling Angiotensin II Type 2 Receptor with ab324679 at 1/100 (5.0 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : no staining on rat liver.
The section was incubated with ab324679 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 citrate buffer (pH 9.0 epitope retrieval solution 2) for 20 mins

• IHC

Supplier Data

Immunohistochemistry - Anti-Angiotensin II Type 2 Receptor antibody [EPR30232-537] - BSA and Azide free (AB326595)

This data was developed using ab324679, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Angiotensin II Type 2 Receptor with ab324679 at 1/100 (5.0 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : no staining on mouse liver.
The section was incubated with ab324679 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 citrate buffer (pH 9.0 epitope retrieval solution 2) for 20 mins