Anti-MAdCAM1 antibody [EPR27223-58] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-MAdCAM1 antibody [EPR27223-58] - BSA and Azide free (AB309488)

This data was developed using ab309487, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) cells labelling MAdCAM1 with ab309487 at 1/50 (10.14 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing cytoplasmic staining in 293T cells transfected with a mouse MADCAM1 expression vector containing a myc tag, but no staining in 293T cells transfected with a human MAdCAM1 expression vector containing a myc tag.Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). is observed. Myc-Tag Mouse mAb (Alexa Fluor® 647) was used to counterstain tubulin at 1/100 0.38ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MAdCAM1 antibody [EPR27223-58] - BSA and Azide free (AB309488)

This data was developed using ab309487, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Rat cardiac muscle tissue labeling MAdCAM1 with ab309487 at 1/100 (5.07 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : No staining on rat cardiac muscle. The section was incubated with ab309487 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-MAdCAM1 antibody [EPR27223-58] - BSA and Azide free (AB309488)

This data was developed using ab309487, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Rat peripheral lymph tissue labeling MAdCAM1 with ab309487 at 1/100 (5.07 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on the high endothelial venules of peripheral lymph nodes in rat jejunum. The section was incubated with ab309487 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-MAdCAM1 antibody [EPR27223-58] - BSA and Azide free (AB309488)

This data was developed using ab309487, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Mouse peripheral lym tissue labeling MAdCAM1 with ab309487 at 1/100 (5.07 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on the high endothelial venules of peripheral lymph nodes in mouse jejunum. The section was incubated with ab309487 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

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-MAdCAM1 antibody [EPR27223-58] - BSA and Azide free (AB309488)

This data was developed using ab309487, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized bEnd.3 (mouse brain endothelioma endothelialpolyoma middle T antigen transformed) cells labelling MAdCAM1 with ab309487 at 1/50 (10.14 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing membranous staining after the treatment with 20 ng/ml TNF alpha for 24 hours in bEnd.3 cells.Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-MAdCAM1 antibody [EPR27223-58] - BSA and Azide free (AB309488)

This data was developed using ab309487, the same antibody clone in a different buffer formulation.

Immunohistochemistry analysis of (Formalin/PFA-fixed paraffin-embedded sections) Rat lymph node tissue labelling MAdCAM1 with ab309487 at 1 : 100 dilution (B), CD3 epsilon with ab237721at 1 : 2000 dilution (C) and CD20 with ab64088 at 1 : 100 dilution (D). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A : merged staining of anti-MAdCAM (green; Opal™520), anti-CD3 (gray; Opal™690) and anti-CD20 (magenta; Opal™570) on rat lymph node.

Panel B : anti-MAdCAM staining high endothelial venules in rat lymph node.

Panel C : ant-CD3 staining B lymphocytes in rat lymph node.

Panel D : ant-CD20 staining B lymphocytes in rat lymph node.

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

The section was incubated in three rounds of staining : in the order of ab309487, ab237721 and ab64088 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MAdCAM1 antibody [EPR27223-58] - BSA and Azide free (AB309488)

This data was developed using ab309487, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling MAdCAM1 with ab309487 at 1/100 (5.07 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : No staining on mouse cardiac muscle. The section was incubated with ab309487 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

• mIHC

Lab

Multiplex immunohistochemistry - Anti-MAdCAM1 antibody [EPR27223-58] - BSA and Azide free (AB309488)

This data was developed using ab309487, the same antibody clone in a different buffer formulation.

Immunohistochemistry analysis of (Formalin/PFA-fixed paraffin-embedded sections) Mouse lymph node tissue labelling MAdCAM1 with ab309487 at 1 : 100 dilution (B), CD3 epsilon with ab237721at 1 : 2000 dilution (C) and CD20 with ab64088 at 1 : 100 dilution (D). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A : merged staining of anti-MAdCAM (green; Opal™520), anti-CD3 (gray; Opal™690) and anti-CD20 (magenta; Opal™570) on mouse lymph node.

Panel B : anti-MAdCAM staining high endothelial venules in mouse lymph node.

Panel C : ant-CD3 staining B lymphocytes in mouse lymph node.

Panel D : ant-CD20 staining B lymphocytes in mouse lymph node.

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

The section was incubated in three rounds of staining : in the order of ab309487, ab237721 and ab64088 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.