The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
应用说明ICC/IF: Use at a concentration of 10 µg/ml.
WB: Use at a concentration of 1 µg/ml. Detects a band of approximately 35 kDa (predicted molecular weight: 30 kDa).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
功能Non-catalytic subunit of AMP-activated protein kinase (AMPK), an energy sensor protein kinase that plays a key role in regulating cellular energy metabolism. In response to reduction of intracellular ATP levels, AMPK activates energy-producing pathways and inhibits energy-consuming processes: inhibits protein, carbohydrate and lipid biosynthesis, as well as cell growth and proliferation. AMPK acts via direct phosphorylation of metabolic enzymes, and by longer-term effects via phosphorylation of transcription regulators. Also acts as a regulator of cellular polarity by remodeling the actin cytoskeleton; probably by indirectly activating myosin. Beta non-catalytic subunit acts as a scaffold on which the AMPK complex assembles, via its C-terminus that bridges alpha (PRKAA1 or PRKAA2) and gamma subunits (PRKAG1, PRKAG2 or PRKAG3).
序列相似性Belongs to the 5'-AMP-activated protein kinase beta subunit family.
结构域The glycogen-binding domain may target AMPK to glycogen so that other factors like glycogen-bound debranching enzyme or protein phosphatases can directly affect AMPK activity.
翻译后修饰Phosphorylated when associated with the catalytic subunit (PRKAA1 or PRKAA2). Phosphorylated by ULK1; leading to negatively regulate AMPK activity and suggesting the existence of a regulatory feedback loop between ULK1 and AMPK.
Lanes 1 - 4: Merged signal (red and green). Green - ab79885 observed at 38 kDa. Red - loading control, ab18058, observed at 124 kDa.
ab79885 was shown to specifically react with AMPK beta 1 when AMPK beta 1 knockout samples were used. Wild-type and AMPK beta 1 knockout samples were subjected to SDS-PAGE. ab79885 at a concentration of 1ug/mL and ab18058 (loading control to Vinculin) at a dilution of 1/10000 were incubated overnight at 4°C. Blots were developed with goat anti-rabbit IgG (H + L) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
Western blot - AMPK beta 1 antibody (ab79885)
All lanes : Anti-AMPK beta 1 antibody (ab79885) 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 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution Developed using the ECL technique
Exposure time : 2 minutesThe 35 kDa band observed is comparable to the molecular weight seen with other commercially available antibodies to human 5'-AMP-activated protein kinase subunit beta-1 (AMPK beta 1).
ICC/IF image of ab79885 stained HeLa 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 (ab79885, 10µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabbit IgG (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. This antibody also gave a positive result in 4% PFA fixed (10 min) HepG2, and MCF-7 cells at 10µg/ml, and in 100% Methanol fixed (5 min) HeLa, HepG2, MCF-7 cells at 10µg/ml.
Anti-AMPK beta 1 antibody (ab79885)参考文献
has not yet been referenced specifically in any publications.