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Synthetic peptide conjugated to KLH derived from within residues 550 to the C-terminus of Human Lamin A.
(Peptide available as ab27812.)
Our Abpromise guarantee covers the use of ab26300 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 1 µg/ml.|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 76 kDa (predicted molecular weight: 74 kDa).|
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: Lamin A knockout HAP1 cell lysate (20 µg)
Lane 3: A431 cell lysate (20 µg)
Lane 4: NIH3T3 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab26300 observed at 76 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab26300 was shown to recognize Lamin A when Lamin A knockout samples were used, along with additional cross-reactive bands. Wild-type and Lamin A knockout samples were subjected to SDS-PAGE. ab26300 1ug/ml and ab8245 (loading control to GAPDH) at a dilution of 1/1000 were incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
ab26300 stained in Hela cells. Cells were fixed with 100% methanol (5 min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab26300 at 1µg/ml and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control) at 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150120 (pseudo-colored red) and ab150081 (colored green) used at 1 ug/ml for 1hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43µM for 1hour at room temperature.
Immunocytochemistry/ Immunofluorescence analysis of hESCs labeling Lamin A with ab26300 at 1/500 dilution. Samples were fixed with 3.7% paraformaldehyde for 1 hour, and stained for nuclear DNA (DAPI), filamentous actin, tumor recognition antigen 1–81, and nuclear envelope protein Lamin A. For staining, cells were permeabilized with 0.1% Triton X-100 for 10 min. Goat serum, 10%, in phosphate-buffered saline was used to block nonspecific binding for 20 min.
This image is courtesy of an anonymous abreview.
Blocking: 5% milk for 30 minutes at 22°C
Immunocytochemistry/ Immunofluorescence analysis of human vascular smooth muscle cell labeling Lamin A with ab26300 at 1/200 dilution. Cells were fixed in formaldehyde and permeabilized with np40. Cells were blocked with 3% BSA for 1 hour at 21°C. A polyclonal donkey anti-rabbit Alexa Fluor® 568 conjugated secondary antibody was used at 1/500 dilution.
This image is courtesy of an anonymous Abreview
Immunocytochemistry/ Immunofluorescence analysis of Human Lung Fibroblasts labeling Lamin A with ab26300 at 1/500 dilution. Samples were fixed with 3.7% paraformaldehyde for 1 hour, and stained for nuclear DNA (DAPI), filamentous actin, tumor recognition antigen 1–81, and nuclear envelope protein Lamin A. For staining, cells were permeabilized with 0.1% Triton X-100 for 10 min. Goat serum, 10%, in phosphate-buffered saline was used to block nonspecific binding for 20 min.
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