Anti-PAK4 + PAK5 + PAK6 (phospho S474 + S560 + S602) antibody [EPR2471(2)] - BSA and Azide free

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-PAK4 + PAK5 + PAK6 (phospho S474 + S560 + S602) antibody [EPR2471(2)] - BSA and Azide free (AB248680)

This data was developed using ab134097, the same antibody clone in a different buffer formulation.

Overlay histogram showing Hela cells stained with ab134097 (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 (ab134097, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1µg/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 antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

• WB

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Western blot - Anti-PAK4 + PAK5 + PAK6 (phospho S474 + S560 + S602) antibody [EPR2471(2)] - BSA and Azide free (AB248680)

This data was developed using ab134097, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-PAK4 + PAK5 + PAK6 (phospho S474 + S560 + S602) antibody [EPR2471(2)] (<a href='/en-us/products/primary-antibodies/pak4-pak5-pak6-phospho-s474-s560-s602-antibody-epr24712-ab134097'>ab134097</a>) at 1/1000 dilution

Lane 1:

HeLa cell lysate untreated at 10 µg

Lane 2:

HeLa cell lysate treated with Lambda Phosphatase at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 64 kDa

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• OI-RD Scanning

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OI-RD Scanning - Anti-PAK4 + PAK5 + PAK6 (phospho S474 + S560 + S602) antibody [EPR2471(2)] - BSA and Azide free (AB248680)

We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody. Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.