All lanes : Anti-Factor H antibody [OX-24] (ab118820) at 1 µg/ml

Lane 1 : Human serum diluted 1/1000 at 10 µl
Lane 2 : Human plasma diluted 1/1000 at 10 µl
Lane 3 : Purified Factor H protein at 0.5 µg
Lane 4 : Purified Factor H protein at 0.1 µg

Performed under reducing conditions.

Predicted band size: 139 kDa
Observed band size: 170 kDa why is the actual band size different from the predicted?

This blot was produced using a 4-12% Bis-tris under the MOPS buffer system. The gel was run at 200V for 55 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab118820 overnight at 4°C at a 1ug/ml concentration. Antibody binding was detected using Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) at 1/20000 dilution for 1 hour at room temperature before imaging.

This image was generated using the ascites version of the product.

96-well microtitre plates were coated overnight at 4°C with recombinant human C3, C3b, iC3b, and Factor H proteins, in duplicate at a concentration of 1µg/mL. Plates were blocked with 1% BSA in PBS-T (0.1% Tween®) for 1 hour before incubation with a 10-step 4x serial dilution of ab118820 from 10µg/mL for 1 hour at room temperature. Antibody binding was detected with Goat Anti-Mouse IgG H&L (HRP) (ab6789) secondary antibody at a 1 in 10000 dilution for 1 hour at room temperature. Plates were incubated with TMB ELISA substrate for 7 minutes prior to being stopped with Stop solution and absorbance measured at 450nm.