Western blot

All lanes : Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048) at 1/1000 dilution

Lane 1 : Hela whole cell lysate
Lane 2 : Hela whole cell lysate with Lamin B1 peptide (ab16375) at 1 µg/ml

Lysates/proteins at 20 µg per lane.

Secondary
Alexa fluor goat polyclonal to Rabbit IgG at 1/10000 dilution

Performed under reducing conditions.

Predicted band size : 66 kDa
Observed band size : 68-70 kDa (why is the actual band size different from the predicted?)

Immunocytochemistry/ Immunofluorescence - Lamin B1 antibody - Nuclear Envelope Marker (ab16048)

ICC/IF image of ab16048 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab16048, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iT^TM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).

Immunocytochemistry - Lamin B1 antibody - Nuclear Envelope Marker (ab16048)

Human and mouse cells stained with ab16048 (1/500). The cells were fixed in 100% methanol for 6 minutes at -20°C, then washed once in PBS.

A: HeLa cells + ab16048 (green)
B: HeLa cells counterstained with DAPI (blue)
C: 3T3 cells + ab16048 (green)
D: 3T3 cells counterstained with DAPI (blue)

Image courtesy of Marilena Ciciarello and Patrizia Lavia, University 'La Sapienza' CNR, Italy

Immunocytochemistry - Lamin B1 antibody - Nuclear Envelope Marker (ab16048)

Human and mouse cells stained with ab16048 (1/500). The cells were fixed and permeabilized in 4% formaldehyde, 0.2% Tritonm X100 for 10 minutes at room temperature, then washed 3x in PBS.

A: HeLa cells + ab16048 (green)
B: HeLa cells counterstained with DAPI (blue)
C: 3T3 cells + ab16048 (green)
D: 3T3 cells counterstained with DAPI (blue)

Image courtesy of Marilena Ciciarello and Patrizia Lavia, University 'La Sapienza' CNR, Italy

Western blot - Lamin B1 antibody - Nuclear Envelope Marker (ab16048)

Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048) at 1/1000 dilution + Pancreatic cell line - whole cell lysate at 20 µg

Secondary
HRP conjugated goat anti-rabbit antibody at 1/2000 dilution
developed using the ECL technique

Performed under reducing conditions.

Predicted band size : 66 kDa
Observed band size : 68 kDa (why is the actual band size different from the predicted?)


Exposure time : 30 seconds

This image is courtesy of an anonymous Abreview

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Lamin B1 antibody - Nuclear Envelope Marker (ab16048)

IHC image of Lamin B1 staining in human liver FFPE section, performed on a Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab16048, 1µg/ml, for 8 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.