Anti-ATP-binding cassette sub-family A member 3 antibody [EPR29622-109]

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-ATP-binding cassette sub-family A member 3 antibody [EPR29622-109] (AB324911)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse lung tissue staining ATP-binding cassette sub-family A member 3 with ab324911 at a 1 : 2000 (0.253 ug/ml) dilution, ab322404 anti-PSAP used at 1 : 5000 (0.092 ug/ml) dilution and ab305303 anti-Aquaporin-5 used at a 1 : 2000 (0.265 ug/ml) dilution.

Panel A : merged staining of anti-ATP-binding cassette sub-family A member 3 (green; Opal™690), anti-PSAP (magenta; Opal™520) and anti-Aquaporin-5 (yellow; Opal™570) on mouse lung.
Panel B : anti-ATP-binding cassette sub-family A member 3 staining typeⅡ pneumocytes in mouse lung.
Panel C : ant-PSAP staining typeⅡ pneumocytes in mouse lung.
Panel D : ant-Aquaporin-5 staining typeⅠ pneumocytes in mouse lung.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab324911, ab322404 and ab305303 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.

Nuclear counter stain with DAPI.

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-ATP-binding cassette sub-family A member 3 antibody [EPR29622-109] (AB324911)

Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling ATP-binding cassette sub-family A member 3 with ab324911 at 1/2000 (0.253 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : No staining on mouse cardiac muscle (PMID : 17142808).
The primary antibody was incubated 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-ATP-binding cassette sub-family A member 3 antibody [EPR29622-109] (AB324911)

Immunohistochemical analysis of paraffin-embedded Mouse lung adenocarcinoma tissue labeling ATP-binding cassette sub-family A member 3 with ab324911 at 1/2000 (0.253 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse lung adenocarcinoma.
The primary antibody was incubated 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-ATP-binding cassette sub-family A member 3 antibody [EPR29622-109] (AB324911)

Immunohistochemical analysis of paraffin-embedded Mouse cerebellum tissue labeling ATP-binding cassette sub-family A member 3 with ab324911 at 1/2000 (0.253 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on Purkinje cells in mouse cerebellum.
The primary antibody was incubated 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-ATP-binding cassette sub-family A member 3 antibody [EPR29622-109] (AB324911)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling ATP-binding cassette sub-family A member 3 with ab324911 at 1/2000 (0.253 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse kidney tubules.
The primary antibody was incubated 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-ATP-binding cassette sub-family A member 3 antibody [EPR29622-109] (AB324911)

Immunohistochemical analysis of paraffin-embedded Mouse lung tissue labeling ATP-binding cassette sub-family A member 3 with ab324911 at 1/2000 (0.253 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on typeⅡ pneumocytes in mouse lung.
The primary antibody was incubated 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

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Western blot - Anti-ATP-binding cassette sub-family A member 3 antibody [EPR29622-109] (AB324911)

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

Low expression : heart, skeletal muscle (PMID : 17142808), C2C12.

The molecular weight observed is consistent with what have been described in literatures (PMID : 17142808; PMID : 20863830).

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

The identity of the lower MW bands below 150 kDa are unknown.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-ATP-binding cassette sub-family A member 3 antibody [EPR29622-109] (ab324911) at 1/1000 dilution

Lane 1:

Mouse lung tissue lysate at 20 µg

Lane 2:

Mouse kidney tissue lysate at 20 µg

Lane 3:

Mouse heart tissue lysate at 20 µg

Lane 4:

Mouse skeletal muscle tissue lysate at 20 µg

Lane 5:

Mouse liver tissue lysate at 20 µg

Lane 6:

Mouse spleen tissue lysate at 20 µg

Lane 6:

Hepa1-6 (mouse hepatoma epithelial cell) whole cell lysate at 20 µg

Lane 8:

C2C12 (mouse myoblast) 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 1/20000 dilution

Observed band size: 150 kDa,190 kDa,124 kDa

false

Exposure time: 37s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATP-binding cassette sub-family A member 3 antibody [EPR29622-109] (AB324911)

Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling ATP-binding cassette sub-family A member 3 with ab324911 at 1/2000 (0.253 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : No staining on mouse skeletal muscle (PMID : 17142808).
The primary antibody was incubated 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

• Dot

Supplier Data

Dot Blot - Anti-ATP-binding cassette sub-family A member 3 antibody [EPR29622-109] (AB324911)

Dot blot analysis of ATP-binding cassette sub-family A member 3 using ab324911 at 1 : 1000 (0.505 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Lane1 : His-tagged mouse ATP-binding cassette sub-family A member 3 recombinant protein 100 ng
Lane2 : His-tagged mouse Phospholipid-transporting ATPase ABCA17 recombinant protein 100 ng
Lane3 : His-tagged mouse Phospholipid-transporting ATPase ABCA4 recombinant protein 200 ng
Lane4 : His-tagged mouse Phospholipid-transporting ATPase ABCA2 recombinant protein 500 ng

Exposure time : 26 seconds.

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

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

All lanes:

Dot Blot - Anti-ATP-binding cassette sub-family A member 3 antibody [EPR29622-109] (ab324911) at 1/1000 dilution

Secondary

All lanes:

Dot 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

false