Anti-Aquaporin 3 antibody [EPR19932] - BSA and Azide free

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Aquaporin 3 antibody [EPR19932] - BSA and Azide free (AB240393)

This data was developed using ab215853, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human skin tissue staining Filaggrin with ab221155 at a 1/1000 dilution, ab215853 anti-Aquaporin 3 used at an appropriate dilution and ab321976 anti-LAMA3 used at an appropriate dilution.

Panel A : merged staining of anti-Filaggrin (green; Opal^™520), anti-Aquaporin 3 (magenta; Opal^™690) and anti-LAMA3 (gray; Opal^™570) on human skin.

Panel B : anti-Filaggrin staining maturing squamous cells in human skin.

Panel C : anti-Aquaporin 3 staining membrane of squamous cells in human skin.

Panel D : anti-LAMA3 staining epithelial basement membrane in human skin.

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

The section was incubated in three rounds of staining : in the order of ab221155, ab215853 and ab321976 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 using Opal Polymer HRP Ms + Rb secondary reagent. 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aquaporin 3 antibody [EPR19932] - BSA and Azide free (AB240393)

Immunohistochemical analysis of paraffin-embedded human skin squamous cell cancer tissue labeling Aquaporin 3 with ab215853 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Membranous staining on human skin squamous cell cancer is observed.

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab215853).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aquaporin 3 antibody [EPR19932] - BSA and Azide free (AB240393)

Immunohistochemical analysis of paraffin-embedded human cervix cancer tissue labeling Aquaporin 3 with ab215853 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Membranous staining on cervix cancer is observed.

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab215853).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aquaporin 3 antibody [EPR19932] - BSA and Azide free (AB240393)

Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling Aquaporin 3 with ab215853 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Membranous staining on kidney collecting ducts is observed.

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab215853).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aquaporin 3 antibody [EPR19932] - BSA and Azide free (AB240393)

Immunohistochemical analysis of paraffin-embedded human bladder cancer tissue labeling Aquaporin 3 with ab215853 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Membranous staining on bladder cancer is observed.

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab215853).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aquaporin 3 antibody [EPR19932] - BSA and Azide free (AB240393)

Immunohistochemical analysis of paraffin-embedded human skin tissue labeling Aquaporin 3 with ab215853 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Membranous staining on skin squamous cell is observed.

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab215853).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Aquaporin 3 antibody [EPR19932] - BSA and Azide free (AB240393)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab215853).

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized A431 (human epidermoid carcinoma epithelial cell) labelling Aquaporin 3 with ab215853 at 1/100 (5.03 μg/ml) dilution (Green). Confocal image showing mainly membranous staining in A431 cell line.

Negative control : U-87 MG

Nuclear DNA was labelled with DAPI (shown in blue). Secondary used was ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 μg/ml). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5μg/ml) dilution (Magenta).

Negative control : U-87 MG

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).