Immunohistochemistry - Anti-Phospho-PAK4/5/6 (Ser474/Ser560/Ser602) Rabbit mAb [24P90Y33]

Sample: Paraformaldehyde-fixed, paraffin embedded rat kidney tissue

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

Primary antibody: R015436 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-PAK4/5/6 (Ser474/Ser560/Ser602) Rabbit mAb [24P90Y33]

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

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

Primary antibody: R015436 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-PAK4/5/6 (Ser474/Ser560/Ser602) Rabbit mAb [24P90Y33]

Sample: Paraformaldehyde-fixed, paraffin embedded mouse liver tissue

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

Primary antibody: R015436 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-PAK4/5/6 (Ser474/Ser560/Ser602) Rabbit mAb [24P90Y33]

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: R015436 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-Phospho-PAK4/5/6 (Ser474/Ser560/Ser602) Rabbit mAb [24P90Y33]

All lanes: R015436 at 1:1,000 dilution

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

Lane 2: Huh1 (Human hepatocarcinoma epithelial cell) whole cell lysates

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

Lane 4: T24 (Human bladder cancer epithelial cell) whole cell lysates

Lane 5: U87 (Human malignant glioblastoma epithelial cell) whole cell lysates

Lane 6: Mouse brain whole tissue lysates

Lane 7: PC-12 (Rat adrenal pheochromocytoma epithelial cell) whole cell lysates

Lane 8: Rat heart 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: 80 kDa

Observed band size: 80 kDa

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