Immunofluorescent analysis of beta-actin (green) in HeLa cells. Cells were fixed with 4% paraformaldehyde (PFA) for 15 minutes at room temperature and permeabilized with 0.5% Triton X-100 in PBS for 15 minutes at room temperature (left panel) or fixed and permeabilized with ice cold methanol for 10 minutes at room temperature (middle panel). Cells were blocked with 0.3% BSA in PBS for 15 minutes at room temperature, and probed with a beta-actin polyclonal antibody (Product # PA1-183) at a dilution of 1:100 for at least 1 hour at room temperature. Cells were then washed with PBS, and incubated with a DyLight 488 goat anti-rabbit IgG secondary antibody (Product # 35552) at a dilution of 1:500 for 30 minutes at room temperature. As a positive control, PFA-fixed cells were stained with DyLight 488-conjugated Phalloidin (Product # 21833) at a dilution of 1:1000 (right panel). Nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification.
|Tested species reactivity||Human, Mouse, Primate, Rat|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Purified full-length recombinant human beta-actin expressed in E. coli|
|Storage buffer||PBS with 30% glycerol, 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunofluorescence (IF)||1:50 - 1:200|
|Immunoprecipitation (IP)||2 µg|
|Western Blot (WB)||1:500 - 1:2000|
By Western blot, PA1-183 detects a prominent ~42kD protein in human, mouse, non-human primate, and rat whole cell lysates.
For visualization of actin filaments by immunofluorescence, fixation and permeabilization with methanol is required.
Beta-actin is one of six different actin isoforms which have been identified. Actins are highly conserved proteins that are involved in cell motility, structure and integrity. Because beta-actin is ubiquitously expressed in all eukaryotic cells, it is frequently used as a loading control for assays involving protein detection, such as Western blotting. Antibodies to beta-actin provide a specific and useful tool in studying the intracellular distribution of beta-actin and the static and dynamic aspects of the cytoskeleton.
ACTB; actin, beta; actin, cytoplasmic 1; Atcb; beta cytoskeletal actin; beta-actin; cytoplasmic beta-actin; PS1TP5-binding protein 1
ACTB; Actx; beta-actin; BRWS1; E430023M04Rik; PS1TP5BP1