All lanes : Anti-iNOS antibody [K13-A] (ab136918) at 1/2000 dilution

Lane 1 : Raw264.7 Whole Cell Lysate
Lane 2 : Raw264.7 Whole Cell Lysate (LPS stimulated)

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat anit Rabbit IR680 at 1/10000 dilution

Performed under reducing conditions.

Predicted band size: 131 kDa
Observed band size: 140 kDa why is the actual band size different from the predicted?
Additional bands at: 15 kDa, 36 kDa, 42 kDa, 50 kDa. We are unsure as to the identity of these extra bands.

This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab136918 overnight at 4°C. Antibody binding was detected using Goat anti Mouse IR680 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx

Anti-iNOS antibody [K13-A] (ab136918) at 1/2000 dilution + iNOS Overexpression Lysate (Human) at 0.1 µg

Secondary
Goat anti Rabbit IR680 at 1/10000 dilution

Performed under reducing conditions.

Predicted band size: 131 kDa
Observed band size: 140 kDa why is the actual band size different from the predicted?

This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab136918 overnight at 4°C. Antibody binding was detected using Goat anti Rabbit IR680 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx

Anti-iNOS antibody [K13-A] (ab136918) at 1/2000 dilution + Mouse brain lysate at 50 µg

Predicted band size: 131 kDa