Anti-PAK1 (phospho T212) antibody [EPR20045] - BSA and Azide free

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-PAK1 (phospho T212) antibody [EPR20045] - BSA and Azide free (AB235116)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling PAK1 (phospho T212) with ab223852 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1,000 dilution (green). Confocal image showing positive staining on HeLa cells in M phase. The signal decreased after treatment with Lambda Protein Phosphatase 31°C for 2 hours (treated).

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution (red).

The negative control : PBS followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1,000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab223852).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-PAK1 (phospho T212) antibody [EPR20045] - BSA and Azide free (AB235116)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A549 (human lung carcinoma cell line) cells labeling PAK1 (phospho T212) with ab223852 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1,000 dilution (green). Confocal image showing positive staining on HeLa cells in M phase. The signal decreased after treatment with Lambda Protein Phosphatase 31°C for 2 hours (treated).

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).

The negative control : PBS followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1,000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab223852).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAK1 (phospho T212) antibody [EPR20045] - BSA and Azide free (AB235116)

Immunohistochemical analysis of paraffin-embedded human prostate tissue labeling PAK1 (phospho T212) with ab223852 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Sporadic nuclear staining on human prostate (PMID : 19876919; PMID : 12176334) is observed. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab223852).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAK1 (phospho T212) antibody [EPR20045] - BSA and Azide free (AB235116)

Immunohistochemical analysis of paraffin-embedded human colon cancer tissue labeling PAK1 (phospho T212) with ab223852 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining on tumor cells of human colon cancer (PMID : 19876919; PMID : 12176334) is observed. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab223852).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-PAK1 (phospho T212) antibody [EPR20045] - BSA and Azide free (AB235116)

Intracellular flow cytometric analysis of 80% methanol-fixed, 0.1% Tween-20 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cell line labeling PAK1 (phospho T212) with ab223852 at 1/50 (right panel) compared with a Rabbit monoclonal IgG (ab172730) Isotype control (left panel). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077), at 1/2,000 dilution was used as the secondary antibody. Unlabeled control cells without incubation with primary antibody and secondary antibody (Blue).

Cells were pretreated with 20 μg/ml RNase A for 30 minutes to eliminate the non-specific binding between RNA and PI (Propidium iodide). PAK1 (phospho T212) is highly expressed in mitotic cells (PMID : 12176334).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab223852).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAK1 (phospho T212) antibody [EPR20045] - BSA and Azide free (AB235116)

Immunohistochemical analysis of paraffin-embedded human lung squamous cancer tissue labeling PAK1 (phospho T212) with ab223852 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining on tumor cells of human lung squamous cancer (PMID : 19876919; PMID : 12176334) is observed. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab223852).

• Dot

Supplier Data

Dot Blot - Anti-PAK1 (phospho T212) antibody [EPR20045] - BSA and Azide free (AB235116)

Dot blot analysis of PAK1 (phospho T212) peptide using ab223852 1/1,000 dilution. Goat Anti-Rabbit IgG H&L (HRP) (ab97051), was used as secondary antibody at 1/100,000 dilution.

Lane 1 : PAK1 (phospho T212) peptide.

Lane 2 : PAK1 non-phospho peptide.

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure time : 3 minutes.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab223852).