Anti-Sumo 2 + Sumo 3 + Sumo 4 antibody [EPR7163]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Sumo 2 + Sumo 3 + Sumo 4 antibody [EPR7163] (AB126606)

ab126606 at 1/250 dilution staining Sumo 2 + Sumo 3 + Sumo 4 in HepG2 (Human liver hepatocellular carcinoma cell line) cells by immunofluorescence.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Sumo 2 + Sumo 3 + Sumo 4 antibody [EPR7163] (AB126606)

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.

• WB

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Western blot - Anti-Sumo 2 + Sumo 3 + Sumo 4 antibody [EPR7163] (AB126606)

All lanes:

Western blot - Anti-Sumo 2 + Sumo 3 + Sumo 4 antibody [EPR7163] (ab126606) at 1/10000 dilution

Lane 1:

HepG2 cell lysate at 10 µg

Lane 2:

293T cell lysate at 10 µg

Lane 3:

Jurkat cell lysate at 10 µg

Lane 4:

Human spleen tissue lysate at 10 µg

Secondary

All lanes:

Goat anti-Rabbit HRP at 1/2000 dilution

Predicted band size: 11 kDa

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• WB

Lab

Western blot - Anti-Sumo 2 + Sumo 3 + Sumo 4 antibody [EPR7163] (AB126606)

Blocking buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-Sumo 2 + Sumo 3 + Sumo 4 antibody [EPR7163] (ab126606) 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

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Exposure time: 3s

• OI-RD Scanning

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OI-RD Scanning - Anti-Sumo 2 + Sumo 3 + Sumo 4 antibody [EPR7163] (AB126606)

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.