All lanes : Anti-C3 antibody [EPR2988] (ab181147) at 1/1000 dilution (purified)

Lane 1 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate
Lane 2 : Human plasma lysate
Lane 3 : Human serum lysate
Lane 4 : Human spinal cord lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 187 kDa
Observed band size: 115,34,40 kDa why is the actual band size different from the predicted?

The molecular weights observed are consistent with what have been described in the literature (PMID: 23619360)

Immunocytochemistry/ Immunofluorescence analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling C3 with purified ab181147 at 1/250 dilution (0.4 µg/mL). Cells were fixed in 100% Methanol and permeabilized with None. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1/200 (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

Flow Cytometry analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling C3 with purified ab181147 at 1/20 dilution (5 μg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).