Immunofluorescent analysis of beta-actin (green) in HeLa cells. Cells were fixed and permeabilized with ice-cold methanol for 10 minutes at room temperature, and blocked with 0.3% BSA in PBS for 15 minutes at room temperature. Cells were probed with a beta-actin monoclonal antibody (Product # MA1-140) at a dilution of 1:2000, or incubated in blocking buffer as a negative control, overnight at 4°C, washed with PBS, and incubated with a DyLight 488-conjugated goat anti-mouse IgG secondary antibody (Product # 35502) at a dilution of 1:1000 for 30 minutes at room temperature. Nuclei (blue) were stained with DAPI (Product # 62248). Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification.
|Tested species reactivity||Human, Mouse, Non-human primate, Rat|
|Published species reactivity||Human|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Purified full-length recombinant human beta-actin expressed in E. coli|
|Storage buffer||PBS with 1mg/ml BSA, 30% glycerol|
|Contains||0.05% sodium azide|
|Storage Conditions||Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:5000-1:20,000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 1 publications below|
By Western blot, MA1-140 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.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Proteomic analysis of exosomes derived from human lymphoma cells.
MA1-140 was used in western blot to characterize the proteomic content of lymphoma cell-derived exosomes.
|Yao Y,Wei W,Sun J,Chen L,Deng X,Ma L,Hao S||European journal of medical research (20:null)||2015|