Western blot analysis of beta-Tubulin was performed by loading 75ug of the indicated cell lysates onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a low fluorescence PVDF membrane (Product # 22860) and blocked with SEA BLOCK Blocking Buffer (Product # 37527) for at least 1 hour. The membrane was probed with a DyLight 800-conjugated beta-Tubulin monoclonal antibody (Product # MA5-16308-D800) at a dilution of 1:500 overnight at 4°C on a rocking platform, and washed in TBS-0.1% Tween-20. Fluorescent detection was performed using a near IR imaging system.
|Tested species reactivity||Human, Mouse, Non-human primate, Rat|
|Host / Isotype||Mouse / IgG2a|
|Immunogen||KLH conjugated peptide of the N-terminus of Beta-Tubulin.|
|Conjugate||DyLight 800 4X PEG|
|Storage buffer||PBS with proprietary stabilizer|
|Contains||0.02% sodium azide|
|Storage Conditions||4° C, do not freeze|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:500-1:1000|
* 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.
MA5-16308-D800 detects bet-Tubulin from various species. MA5-16308-D800 has been successfully used for fluorescent detection of beta-Tubulin in Western Blotting.
DyLight 800 4X PEG has an excitation/emission of 783/797 nm.
Microtubules, the major cytoskeletal elements found in all eukaryotic cells, consist of Tubulin, which is a dimer of two 55kDa subunits: alpha and Beta. Microtubules play key roles in chromosome segregation in mitosis, intracellular transport, ciliary and flagellar bending, and structural support of the cytoskeleton. Because beta-tubulin is ubiquitously expressed in all eukaryotic cells, it is frequently used as a loading control for assays involving protein detection, such as Western blotting.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.