Anti-USP28 antibody [EPR4249(2)]

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-USP28 antibody [EPR4249(2)] (AB126604)

Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling USP28 with unpurified ab126604 at 1/200 dilution (1ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488) (ab150077) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) (ab172730) was used as the isotype control, Cell without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-USP28 antibody [EPR4249(2)] (AB126604)

ab126604 at 1/50 dilution staining USP28 in HeLa cells by Immunofluorescence.

• WB

Lab

Western blot - Anti-USP28 antibody [EPR4249(2)] (AB126604)

Western blot : Anti-USP28 antibody [EPR4249(2)] ab126604 staining at 1/1000 dilution, shown in green; Mouse anti-CANX ab238078 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 122 kDa in Wild-type A549 Nuclear cell lysates with no signal observed at this size in USP28 knockout A549 Nuclear cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-USP28 antibody [EPR4249(2)] (ab126604) at 1/1000 dilution

Lane 1:

Wild-type A549 Nuclear cell lysates at 20 µg

Lane 2:

USP28 knockout A549 Nuclear cell lysates at 20 µg

Lane 3:

HeLa cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 122 kDa

Observed band size: 122 kDa

false

• WB

Unknown

Western blot - Anti-USP28 antibody [EPR4249(2)] (AB126604)

All lanes:

Western blot - Anti-USP28 antibody [EPR4249(2)] (ab126604) at 1/1000 dilution

Lane 1:

293T cell lysate at 10 µg

Lane 2:

HeLa cell lysate at 10 µg

Lane 3:

HT1376 cell lysate at 10 µg

Lane 4:

Human heart tissue lysate at 10 µg

Lane 5:

SW480 cell lysate at 10 µg

Lane 6:

A431 cell lysate at 10 µg

Secondary

All lanes:

Goat anti-Rabbit HRP at 1/2000 dilution

Predicted band size: 122 kDa

false

• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-USP28 antibody [EPR4249(2)] (AB126604)

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.