Anti-Lin28 antibody [6D1F9] (ab76369) at 1/2000 dilution + Cell lysates prepared from human NTERA-2 cells

Predicted band size: 23 kDa

Standard Curve for Lin28 (Analyte: Lin28 protein (His tag) (ab89225)); dilution range 1 pg/ml to 1 ug/ml using Capture Antibody Mouse monoclonal [6D1F9] to Lin28 (ab76369) at 0.2 ug/ml and Detector Antibody Rabbit polyclonal to Lin28 (ab63740) at 0.1 ug/ml.

Immunofluorescence staining of methanol fixed HeLa cells were transfected with pMX construct of human Lin28. Cells were analyzed ~62 hours after transfection.

ab76369 at 1/2000 dillution staining Lin28 in NTERA-2 cells by Immunocytochemistry/ Immunofluorescence. The green color in image show positive staining with primary antibody while nuclei were stained blue with DRAQ5 fluorescent DNA dye.

Overlay histogram showing HeLa cells stained with ab76369 (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 (ab76369, 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 result in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween for 20 min used under the same conditions.