Western Blot - Anti-Phospho-Smad3 (Ser423/425) Rabbit mAb [11L61K22]

All lanes: R013553 at 1:1,000 dilution

Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysates

Lane 2: MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysates

Lane 3: A549 (human non-small cell lung cancer epithelial cell) whole cell lysates

Lane 4: SCC-9 (human tongue squamous carcinoma epithelial cell) whole cell lysates

Lane 5: C2C12 (mouse myoblasts epithelial cell) whole cell lysates

Lane 6: U2OS (human osteosarcoma epithelial cell) whole cell lysates

Lysates/proteins at 10 µg per lane.

Secondary antibody: Goat Anti-Rabbit IgG(H+L), HRP Conjugated (Cat. No. LF102) at 1:5,000 dilution

Predicted band size: 55 kDa

Observed band size: 55 kDa

Developed using the ECL technique (Cat. No. SQ201).

Immunohistochemistry - Anti-Phospho-Smad3 (Ser423/425) Rabbit mAb [11L61K22]

Sample: Paraformaldehyde-fixed, paraffin embedded rat kidney tissue

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins.

Primary antibody: R013553 at 1:200 dilution

Secondary antibody: Goat Anti-Rabbit IgG (H+L), HRP conjugated at 1:1,000 dilution

DAB was used as the chromogen.

Counter stained with hematoxylin.

Positive/negative staining were presented.

Only the secondary antibody was used as the negative control.

Immunohistochemistry - Anti-Phospho-Smad3 (Ser423/425) Rabbit mAb [11L61K22]

Sample: Paraformaldehyde-fixed, paraffin embedded human endometrial cancer tissue

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins.

Primary antibody: R013553 at 1:200 dilution

Secondary antibody: Goat Anti-Rabbit IgG (H+L), HRP conjugated at 1:1,000 dilution

DAB was used as the chromogen.

Counter stained with hematoxylin.

Positive/negative staining were presented.

Only the secondary antibody was used as the negative control.

Immunofluorescence - Anti-Phospho-Smad3 (Ser423/425) Rabbit mAb [11L61K22]

Sample: Paraformaldehyde-fixed, paraffin embedded human colorectal carcinoma tissue

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins.

Primary antibody: R013553 at 1:100 dilution

Secondary antibody: Goat anti-Rabbit (488) at 1:1,000 dilution (shown in green)

Nuclei were stained with DAPI (shown in blue).

Immunofluorescence - Anti-Phospho-Smad3 (Ser423/425) Rabbit mAb [11L61K22]

Sample: HeLa cells

The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.5% Triton X-100 for 10 minutes and then blocked with 5% BSA in 0.1% PBS-Tween for 0.5 hours.

Primary antibodies: R013553 at 1:100 dilution and α-tubulin Mouse Monoclonal Antibody (Cat. No. LF209) at 1:100 dilution

Secondary antibodies: Goat anti-Rabbit (488) at 1:1,000 dilution (shown in green) and Goat anti-Mouse (CY3) at 1:1,000 dilution (shown in red)

Nuclei were stained with DAPI (shown in blue).

Western Blot - Anti-Phospho-Smad3 (Ser423/425) Rabbit mAb [11L61K22]

All lanes: R013553 at 1:1,000 dilution

Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates

Lane 2: HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates

Lane 3: HCT116 (Human colorectal carcinoma epithelial cell) whole cell lysates

Lysates/proteins at 10 µg per lane.

Secondary antibody: Goat Anti-Rabbit IgG(H+L), HRP Conjugated (Cat. No. LF102) at 1:5,000 dilution

Predicted band size: 55 kDa

Observed band size: 55 kDa

Developed using the ECL technique (Cat. No. SQ201).