The 30kDa band observed is comparable to the molecular weight seen with other commercially available antibodies to SOCS3.

ICC/IF image of ab16030 stained human HeLa cells. The cells were PFA fixed (10 min) and incubated with the antibody (ab16030, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iT^TM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).

All lanes : Anti-SOCS3 antibody (ab16030) at 1 µg/ml

Lane 1 : Untreated Human peripheral blood mononuclear cell lysate
Lane 2 : Human peripheral blood mononuclear cell lysate (treated with aCD3/28)
Lane 3 : Human peripheral blood mononuclear cell lysate (treated with IFNγ)
Lane 4 : SOCS3 protein

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : HRP-conjugated goat anti-rabbit polyclonal IgG
at 1/2000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 24 kDa
Observed band size: 25 kDa why is the actual band size different from the predicted?


Exposure time: 15 minutes

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