Anti-Sumo 2 + Sumo 3 + Sumo 4 antibody [EPR7163] - BSA and Azide free

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Sumo 2 + Sumo 3 + Sumo 4 antibody [EPR7163] - BSA and Azide free (AB232032)

Overlay histogram showing HEK293 cells stained with ab126606 (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 (ab126606, 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 data was developed using the same antibody clone in a different buffer formulation (ab126606).

• WB

Lab

Western blot - Anti-Sumo 2 + Sumo 3 + Sumo 4 antibody [EPR7163] - BSA and Azide free (AB232032)

Blocking buffer : 5% NFDM/TBST

This data was developed using ab126606, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-Sumo 2 + Sumo 3 + Sumo 4 antibody [EPR7163] (<a href='/en-us/products/primary-antibodies/sumo-2-sumo-3-sumo-4-antibody-epr7163-ab126606'>ab126606</a>) at 1/5000 dilution

Lane 1:

Western blot - Recombinant Human Sumo 1 protein (<a href='/en-us/products/proteins-peptides/recombinant-human-sumo-1-protein-ab140417'>ab140417</a>) at 0.01 µg

Lane 2:

Western blot - Recombinant Human Sumo 2 protein (<a href='/en-us/products/proteins-peptides/recombinant-human-sumo-2-protein-ab140420'>ab140420</a>) at 0.01 µg

Lane 3:

Recombinant Human Sumo 3 protein (<a href='/en-us/products/unavailable/recombinant-human-sumo-3-protein-ab140414'>ab140414</a>) at 0.01 µg

Lane 4:

Western blot - Recombinant Human Sumo 4 protein (<a href='/en-us/products/proteins-peptides/recombinant-human-sumo-4-protein-ab157025'>ab157025</a>) at 0.1 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 11 kDa

Observed band size: 16 kDa

true

Exposure time: 3s

• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-Sumo 2 + Sumo 3 + Sumo 4 antibody [EPR7163] - BSA and Azide free (AB232032)

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.