Immunofluorescent analysis of beta-actin (red) in HeLa cells. Cells were fixed with either 4% paraformaldehyde (PFA) for 15 minutes at room temperature followed by permeabilization 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 (right panel). Cells were blocked with 0.3% BSA in PBS for 15 minutes, and probed with a DyLight 550-conjugated beta-actin monoclonal antibody (Product # MA5-15739-D550) at a dilution of 1:50 overnight at 4C. Nuclei (blue) were stained with DAPI (Product # 62247). Images were taken on a ToxInsight Instrument at 20X magnification.
|Tested species reactivity||Chicken, Human, Mouse, Rabbit, Rat|
|Host / Isotype||Mouse / IgG2b|
|Immunogen||Beta-actin N-terminal peptide|
|Storage buffer||PBS with proprietary stabilizer|
|Contains||0.02% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
* 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.
MA5-15739-D550 has been successfully used for Immunofluorescence. DyLight 550 has an excitation/emission of 562/576nm.
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.