Anti-CD105 抗体 [MEM-226] (ab2529)

Standard Curve for CD105 (Analyte: CD105 protein (ab54338)); dilution range 1pg/ml to 1ug/ml using Capture Antibody Mouse monoclonal [MEM-226] to CD105 (ab2529) at 5ug/ml and Detector Antibody Rabbit polyclonal to CD105 (ab21224) at 0.5ug/ml.

Anti-CD105 antibody [MEM-226] (ab2529) at 5 µg/ml + Human colon tissue lysate - total protein (ab30051) at 10 µg

Secondary
Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Observed band size: 80 kDa (why is the actual band size different from the predicted?)
Additional bands at: 45 kDa, 55 kDa. We are unsure as to the identity of these extra bands.

Overlay histogram showing U937 cells stained with ab2529 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab2529, 1µg/1x10⁶ cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in U937 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.