Confocal immunofluorescent analysis of HT-29 cells using Lamin B1 (119D5-F1) Mouse mAb (green) and β-Actin (13E5) Rabbit mAb (Alexa Fluor® 647 Conjugate) #8584 (red). Blue pseudocolor = Propidium Iodide (PI)/RNase Staining Solution #4087 (fluorescent DNA dye).
Aliquot into single-use vials to avoid multiple freeze/thaw cycles and store at -20°C. A slight precipitate may be present, but will not interfere with antibody performance.
Supplied at 1 mg/mL in PBS containing 0.09% sodium azide. Store at -20°C. The product will freeze at -20°C.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalently purified water.
Recommended Fluorochrome-conjugated Anti-Mouse secondary antibodies:
NOTE: Cells should be grown, treated, fixed and stained directly in multi-well plates, chamber slides or on coverslips.
NOTE: All subsequent incubations should be carried out at room temperature unless otherwise noted in a humid light-tight box or covered dish/plate to prevent drying and fluorochrome fading.
posted December 2010
Protocol Id: 145
Lamin B1 (119D5-F1) Mouse mAb recognizes endogenous levels of total Lamin B1 by immunofluroescence. Testing by immunofluorescence was validated for human cells. Reactivity is expected in other species, including mouse and rat.Species Reactivity:
Monoclonal antibody is produced by immunizing animals with purified rat liver lamins.
Lamins are nuclear membrane structural components that are important in maintaining normal cell functions, such as cell cycle control, DNA replication, and chromatin organization (1-3). Lamins have been subdivided into types A and B. Type-A lamins consist of lamin A and C, which arise from alternative splicing of the lamin A gene LMNA. Lamin A and C are cleaved by caspases into large (41-50 kDa) and small (28 kDa) fragments, which can be used as markers for apoptosis (4,5). Type-B lamins consist of lamin B1 and B2, encoded by separate genes (6-8). Lamin B1 is also cleaved by caspases during apoptosis (9). Research studies have shown that duplication of the lamin B1 gene LMNB1 is correlated with pathogenesis of the neurological disorder adult-onset leukodystrophy (10).
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