Anti-PAK1+PAK2+PAK3 (phospho S141 + S144 + S154) antibody [EP656Y] - BSA and Azide free

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-PAK1+PAK2+PAK3 (phospho S141 + S144 + S154) antibody [EP656Y] - BSA and Azide free (AB239830)

Overlay histogram showing HeLa (Human epithelial cell line from cervix adenocarcinoma) cells stained with unpurified ab40795 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab40795, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight^® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40795).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-PAK1+PAK2+PAK3 (phospho S141 + S144 + S154) antibody [EP656Y] - BSA and Azide free (AB239830)

ab40795 staining PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154) in HeLa (human cervix adenocarcinoma) cells, treated and untreated with Lambda Protein Phosphtase 31°C for 5h by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody. ab7291 and ab150120 were used as counterstains for primary antibody ab75748 and secondary antibody ab150077 respectively and DAPI was used as a nuclear counterstain.

Negative control 1 : Rabbit primary antibody and anti-mouse secondary antibody (ab150120)
Negative control 2 : Mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab150077)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40795).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAK1+PAK2+PAK3 (phospho S141 + S144 + S154) antibody [EP656Y] - BSA and Azide free (AB239830)

ab40795 staining PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154) in human liver carcinoma tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

Negative control 1 : PBS in place of primary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40795).

• IP

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Immunoprecipitation - Anti-PAK1+PAK2+PAK3 (phospho S141 + S144 + S154) antibody [EP656Y] - BSA and Azide free (AB239830)

ab40795 immunoprecipitating PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154). 10μg of HeLa (human cervix adenocarcinoma) whole cell lysate was incubated with primary antibody at a dilution of 1/40 and VeriBlot for IP Detection Reagent (HRP) (ab131366) at a dilution of 1/10000.

Lane 1 : HeLa whole cell lysate (10ug)
Lane 2 : ab40795 IP in HeLa whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab40795 in HeLa (human cervix adenocarcinoma) whole cell lysate

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40795).

All lanes:

Immunoprecipitation - Anti-PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154) antibody [EP656Y] (<a href='/ko/products/primary-antibodies/pak1-phospho-s144-pak2-phospho-s141-pak3-phospho-s154-antibody-ep656y-ab40795'>ab40795</a>)

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• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAK1+PAK2+PAK3 (phospho S141 + S144 + S154) antibody [EP656Y] - BSA and Azide free (AB239830)

ab40795 staining PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154) in rat cerebral cortex tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

Negative control 1 : PBS in place of primary

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40795).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-PAK1+PAK2+PAK3 (phospho S141 + S144 + S154) antibody [EP656Y] - BSA and Azide free (AB239830)

ab40795 staining PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154) in the mousecell line NIH/3T3 (mouse embryo) by intracellular flow cytometry. Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/120. A goat anti rabbit IgG (Alexa Fluor^® 488) at a dilution of 1/500 was used as the secondary antibody.

Isoytype control : Rabbit monoclonal IgG (Black)

Unlabelled control : Cell without incubation with primary antibody and secondary antibody (Blue)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40795).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAK1+PAK2+PAK3 (phospho S141 + S144 + S154) antibody [EP656Y] - BSA and Azide free (AB239830)

ab40795 staining PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S154) in mouse cerebral cortex tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

Negative control 1 : PBS in place of primary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40795).