Anti-MAdCAM1 antibody [EPR27555-87] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MAdCAM1 antibody [EPR27555-87] - BSA and Azide free (AB307735)

This data was developed using ab307734, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded human skeletal muscle tissue labeling MAdCAM1 with ab307734 at 1/500 dilution (1.0 µg/ml) followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : no staining on human skeletal muscle (PMID : 8989586). The section was incubated with ab307734 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 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 [EPR27555-87] - BSA and Azide free (AB307735)

This data was developed using ab307734, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded human gastric adenocarcinoma tissue labeling MAdCAM1 with ab307734 at 1/500 dilution (1.0 µg/ml) followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on blood vessels of human gastric adenocarcinoma. The section was incubated with ab307734 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 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 [EPR27555-87] - BSA and Azide free (AB307735)

This data was developed using ab307734, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded human stomach tissue labeling MAdCAM1 with ab307734 at 1/500 dilution (1.0 µg/ml) followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on blood vessels of human stomach (PMID : 14742547). The section was incubated with ab307734 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 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 [EPR27555-87] - BSA and Azide free (AB307735)

This data was developed using ab307734, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded cell pellets labeling MAdCAM1 with ab307734 at 1/2000 dilution (0.25 µg/ml) followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on HEK-293T transfected with a Myc-His-tagged MADCAM1 construct (image A). No staining on HEK-293T transfected with empty plasmid (image B). The section was incubated with ab307734 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 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 [EPR27555-87] - BSA and Azide free (AB307735)

This data was developed using ab307734, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded human colon tissue labeling MAdCAM1 with ab307734 at 1/500 dilution (1.0 µg/ml) followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on blood vessels of human colon (PMID : 8989586). The section was incubated with ab307734 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 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.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-MAdCAM1 antibody [EPR27555-87] - BSA and Azide free (AB307735)

This data was developed using ab307734, the same antibody clone in a different buffer formulation. Flow cytometric analysis of 2% paraformaldehyde fixed 0.1% Tween-20 permeabilized HEK-293T (human embryonic kidney) cells transfected with a human MAdCAM1 expression vector containing a myc-his tag cells labelling MAdCAM1 with ab307734 at 1/500 dilution (0.1ug)/ Right (Red) compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Cells were surface stained with ab307734, then fixed with 2% PFA followed by intracellular staining with Myc-tag conjugated to Alexa Fluor® 647. Please note Flow Cyt was only tested with over-expression cells.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-MAdCAM1 antibody [EPR27555-87] - BSA and Azide free (AB307735)

This data was developed using ab307734, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293T (human embryonic kidney epithelial cell) cells labeling MAdCAM1 with ab307734 at 1/1000 dilution (0.5 µg/ml) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (2 µg/ml) (Green). Confocal image showing positive staining in HEK-293T cells transfected with a human MAdCAM1 expression vector containing a myc tag. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Myc-Tag Mouse mAb (Alexa Fluor® 647) was used as a counterstain at 1/100 dilution (0.38 µg/ml) (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 dilution (2 µg/ml).

• WB

Lab

Western blot - Anti-MAdCAM1 antibody [EPR27555-87] - BSA and Azide free (AB307735)

This data was developed using ab307734, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST This antibody reacts with human MAdCAM1 but does not react with mouse MAdCAM1. In Western blot, anti-His antibody (ab213204) staining at 1/5000 dilution. Exposure time : 180 seconds

All lanes:

Western blot - Anti-MAdCAM1 antibody [EPR27555-87] (<a href='/en-us/products/primary-antibodies/madcam1-antibody-epr27555-87-ab307734'>ab307734</a>) at 1/1000 dilution

Lane 1:

His-tagged human MAdCAM1 recombinant fragment at 10 ng

Lane 2:

293T cells transfected with a mouse MAdCAM1 expression vector containing a his tag 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 1/100000 dilution

Observed band size: 32 kDa

false

Exposure time: 180s