Immunofluorescence - Anti-Smad2/3 Rabbit mAb [14L86C22]

Sample: Hela cells

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

Primary antibodies: R013762 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 (555) at 1:1,000 dilution (shown in red)

Nuclei were stained with DAPI (shown in blue).

Immunofluorescence - Anti-Smad2/3 Rabbit mAb [14L86C22]

Sample: A431 cells

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

Primary antibodies: R013762 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 (555) at 1:1,000 dilution (shown in red)

Nuclei were stained with DAPI (shown in blue).

Western Blot - Anti-Smad2/3 Rabbit mAb [14L86C22]

All lanes: R013762 at 1:4,000 dilution

Lane 1: C2C12 (mouse myoblasts epithelial cell) whole cell lysates

Lane 2: Jurkat (human T lymphocytic leukemia cell) whole cell lysates

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

Lane 4: RAW264.7 (mouse mononuclear macrophage leukemia epithelial cell)

Lane 5: HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysates

Lane 6: Rat stomach whole tissue lysates

Lane 7: Balb/c mouse liver whole tissue 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: 52 kDa

Observed band size: 58 kDa, 62 kDa

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

Western Blot - Anti-Smad2/3 Rabbit mAb [14L86C22]

All lanes: R013762 at 1:1,000 dilution

Lane 1: MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysates

Lane 2: T24 (human bladder cancer epithelial cell) whole cell lysates

Lane 3: HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysates

Lane 4: SW620 (human colorectal carcinoma epithelial cell) whole cell lysates

Lane 5: Rat brain whole tissue lysates

Lane 6: Rat liver whole tissue 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: 52 kDa

Observed band size: 58-62 kDa

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

Immunohistochemistry - Anti-Smad2/3 Rabbit mAb [14L86C22]

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

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

Primary antibody: R013762 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-Smad2/3 Rabbit mAb [14L86C22]

Sample: Paraformaldehyde-fixed, paraffin embedded mouse heart tissue

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

Primary antibody: R013762 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-Smad2/3 Rabbit mAb [14L86C22]

Sample: Paraformaldehyde-fixed, paraffin embedded mouse brain tissue

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

Primary antibody: R013762 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.