Anti-Aquaporin 5 antibody [EPR3747] - BSA and Azide free

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-Aquaporin 5 antibody [EPR3747] - BSA and Azide free (AB239904)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human lung tissue staining SFTPA1 with ab325321 at a 1 : 1000 (0.512 ug/ml) dilution, ab213363 anti-CD68 used at 1 : 500 (1.314 ug/ml) dilution and ab239904 anti-Aquaporin 5 used at a 1 : 500 (2.0 ug/ml) dilution.

Panel A : merged staining of anti-SFTPA1 (green; Opal™520), anti-CD68 (magenta; Opal™690) and anti-Aquaporin 5 (gray; Opal™570) on human lung.
Panel B : anti-SFTPA1 staining type 2 pneumonocytes in human lung.
Panel C : anti-CD68 staining macrophages in human lung.
Panel D : anti-Aquaporin 5 staining epithelium and type 1 pneumonocytes in human lung.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab325324, ab213363 and ab239904 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aquaporin 5 antibody [EPR3747] - BSA and Azide free (AB239904)

This data was developed using ab92320, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human stomach tissue sections labeling Aquaporin 5 with purified ab92320 at 1/1200 dilution (0.13 μg/ml). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1/0 dilution. PBS instead of the primary antibody was used as the negative control.

• WB

Lab

Western blot - Anti-Aquaporin 5 antibody [EPR3747] - BSA and Azide free (AB239904)

This data was developed using ab92320 the same antibody clone in a different buffer formulation.

1%SDS Hot lysis method is recommended for this antibody. The molecular weight observed is consistent with what has been described in the literatures (PMID : 32901888, 23313152).

All lanes:

Western blot - Anti-Aquaporin 5 antibody [EPR3747] (<a href='/en-us/products/primary-antibodies/aquaporin-5-antibody-epr3747-ab92320'>ab92320</a>) at 1/1000 dilution

Lane 1:

SW480 (Human colorectal adenocarcinoma epithelial cell) prepared in RIPA lysis method whole cell lysate at 20 µg

Lane 2:

SW480 (Human colorectal adenocarcinoma epithelial cell) prepared in 1%SDS Hot lysis method 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

Predicted band size: 28 kDa

Observed band size: 24 kDa

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