Western blot analysis of Beta-Actin was performed by loading 50 µg of various cell lysates per well onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane and blocked with SEA BLOCK (Product # 37527) for at least 1 hour. The membrane was probed with a DyLight 800-conjugated Beta-Actin monoclonal antibody (Product # MA5-15739-D800) at a dilution of 1:1000 for 1 hour at room temperature on a rocking platform and washed in TBS-0.1% Tween-20. Detection was performed using the LI-COR Odyssey.
|Tested species reactivity||Chicken, Human, Mouse, Rabbit, Rat|
|Host / Isotype||Mouse / IgG2b|
|Immunogen||Beta-actin N-terminal peptide.|
|Conjugate||DyLight 800 4X PEG|
|Storage buffer||PBS with proprietary stabilizer|
|Contains||0.02% sodium azide|
|Storage Conditions||4° C, store in dark|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:500-1:2000|
* 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-D800 has been successfully used for Western blotting and immunofluorescence. DyLight 800 4X PEG has an excitation/emission of 783/797 nm. 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.