Flow cytometry analysis of beta actin in A431 cells (green) compared to an isotype control (blue). Cells were harvested and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 minutes at room temperature and incubated with a beta Actin Loading Control Monoclonal Antibody (BA3R), Biotin (Cat. No. MA5-15739-BTIN). Cells were then incubated for 40 min at room temperature in the dark with a secondary antibody conjugated to DyLight 488 dye, and resuspended in PBS for FACS analysis.
Cell: A431
Concentration: 2ug/test (100ul)
Theory location: Cytoplasm
Beta actin antibodies provide a specific and useful tool for studying the intracellular distribution of beta actin and the static and dynamic aspects of the cytoskeleton. Because it is ubiquitously expressed in all eukaryotic cells, beta actin is frequently used as a loading control. Beta actin is one of six isoforms of actin that have been identified in mammals. Actin is the primary component of the cytoskeleton, and beta actin is predominantly expressed in non-muscle cells, where it controls cell structure and motility. Quality Invitrogen beta actin antibodies are available for a variety of research needs. We also offer fluorescent and enzyme-conjugated versions for your added convenience.
Applications
Click image to enlarge
Immunofluorescence analysis of beta actin was performed using 70% confluent log phase NIH/3T3 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with beta actin Rabbit Polyclonal Antibody (Cat. No. PA1-46296) at 2µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal Secondary Antibody, Alexa Fluor® 488 conjugate (Cat. No. A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade Gold Antifade Mountant with DAPI (Cat. No. S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Cat. No. R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
Western blot analysis of beta actin was performed by loading various cell lysates onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a low-fluorescence PVDF membrane and probed with a beta actin monoclonal antibody conjugated to Alexa Fluor 488 dye (Cat. No. MA1-140-A488).
Resources
For Research Use Only. Not for use in diagnostic procedures.
For Research Use Only. Not for use in diagnostic procedures.