All lanes : Anti-SLC22A3/OCT3 antibody [EPR6630] (ab124826) at 1/10000 dilution

Lane 1 : Human fetal kidney lysate
Lane 2 : Human muscle lysate
Lane 3 : Human placenta lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 61 kDa
Observed band size: 80 kDa why is the actual band size different from the predicted?

Overlay histogram showing JEG3 cells stained with ab124826 (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 (ab124826, 1/100 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.

ab124826, at 1/100 dilution, staining SLC22A3/OCT3 in paraffin-embedded Human kidney tissue by Immunohistochemistry.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Equilibrium disassociation constant (K_D)
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