The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 5 µg/ml.
Use a concentration of 1 µg/ml. Detects a band of approximately 73 kDa (predicted molecular weight: 66 kDa).
機能Lamins are components of the nuclear lamina, a fibrous layer on the nucleoplasmic side of the inner nuclear membrane, which is thought to provide a framework for the nuclear envelope and may also interact with chromatin.
関連疾患Defects in LMNB1 are the cause of leukodystrophy demyelinating autosomal dominant adult-onset (ADLD) [MIM:169500]. ADLD is a slowly progressive and fatal demyelinating leukodystrophy, presenting in the fourth or fifth decade of life. Clinically characterized by early autonomic abnormalities, pyramidal and cerebellar dysfunction, and symmetric demyelination of the CNS. It differs from multiple sclerosis and other demyelinating disorders in that neuropathology shows preservation of oligodendroglia in the presence of subtotal demyelination and lack of astrogliosis.
配列類似性Belongs to the intermediate filament family.
翻訳後修飾B-type lamins undergo a series of modifications, such as farnesylation and phosphorylation. Increased phosphorylation of the lamins occurs before envelope disintegration and probably plays a role in regulating lamin associations.
Lane 1: Wild type HAP1 whole cell lysate (20 µg) Lane 2: Empty Lane 3: LMNB1 whole cell lysate (20 µg) Lane 4: Empty Lanes 1 - 4: Merged signal (red and green). Green - ab90169 observed at 70 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab90169 was shown to specifically react with Lamin B1 when Lamin B1 knockout samples were used. Wild-type and Lamin B1 knockout samples were subjected to SDS-PAGE. ab90169 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10000 dilution respectively. Blots were developed with Donkey anti-Chicken IgG H&L (IRDye® 800CW) and Donkey anti Mouse IgG H&L secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
Western blot - Lamin B1 antibody (ab90169)
All lanes : Anti-Lamin B1 antibody (ab90169) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
Lysates/proteins at 20 µg per lane.
Secondary Goat polyclonal to Chicken IgY - H&L (HRP) at 1/3000 dilution Developed using the ECL technique
ICC/IF image of ab90169 stained HepG2 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab90169 at 5ug overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti- chicken IgY (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.