Anti-Lamin B1 antibody [119D5-F1] - Nuclear Envelope Marker (ab8982)

Immunocytochemistry - Lamin B1 antibody [119D5-F1] - Nuclear Envelope Marker (ab8982)

Methanol fixed HeLa and 3T3 cells were stained with ab8982 (1/50 and 1/20 respectively). The cells were fixed in 100% methanol for 6 minutes at -20°C. ab8982 clearly stains the nuclear envelope with very little backgrouns staining. 

A: HeLa cells + ab8982 (1/50) (green)
B: HeLa cells counterstained with DAPI (blue)
C. 3T3 cells + ab8982 (1/20) (green)
D. 3T3 cells counterstained with DAPI (blue)

Marilena Ciciarello & Patrizia Lavia, University La Sapienza

Flow Cytometry-Lamin B1 antibody [119D5-F1] - Nuclear Envelope Marker(ab8982)

Overlay histogram showing HeLA cells stained with ab8982 (red line). The cells were fixed with 100% 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 (ab8982, 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.

Immunohistochemistry (Frozen sections) - Anti-Lamin B1 antibody [119D5-F1] - Nuclear Envelope Marker (ab8982)

Immunofluorescence staining images of 9 day old zebrafish embryos. Frozen sections fixed in Acetone:Methanol 1:1 ab8982 diluted 1/100, incubated for 45 minutes at room temperature.

Western blot - Anti-Lamin B1 antibody [119D5-F1] - Nuclear Envelope Marker (ab8982)

All lanes : Anti-Lamin B1 antibody [119D5-F1] - Nuclear Envelope Marker (ab8982) at 1/2000 dilution

Lane 1 : Whole cell lysate prepared from human mammary cells (24 hour culture)
Lane 2 : Whole cell lysate prepared from human mammary cells (48 hour culture)

Lysates/proteins at 50000 cells per lane.

Secondary
HRP conjugated goat anti-mouse polyclonal at 1/5000 dilution
developed using the ECL technique

Predicted band size : 67 kDa
Observed band size : 70 kDa (why is the actual band size different from the predicted?)


Exposure time : 40 seconds

Image courtesy of an anonymous Abreview.