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