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Tissue, cells or virus corresponding to Rat Lamin B1 (C terminal). Immunogen = Purified Rat liver lamins
Our Abpromise guarantee covers the use of ab8982 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/100 - 1/1000. Predicted molecular weight: 67 kDa.|
|Dot blot||Use at an assay dependent concentration.|
|ELISA||Use at an assay dependent concentration.|
|Flow Cyt||1/100 - 1/200. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.|
|ICC||1/20 - 1/50.|
|ICC/IF||Use a concentration of 0.5 µg/ml.|
|IHC-Fr||Use at an assay dependent concentration.|
ab8982 staining Lamin B1 in wild-type HAP1 cells (top panel) and LMNB1 knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab8982 at 0.5μg/ml and ab202272 at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Mouse IgG (Alexa Fluor® 488) (ab150117) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Methanol fixed HeLa and 3T3 cells (ab7179) 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)
Image courtesy of an anonymous Abreview.
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 (ab7481) / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab8982, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. 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.
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