Western blot analysis of alpha-Tubulin was performed by loading 20 µg of HCT 116 (lane1), MDA-MB-231 (lane2), Raji (lane3) and A431 (lane4) cell lysate using Novex® NuPAGE® 12 % Bis-Tris gel (NP0341BOX), XCell SureLock™ Electrophoresis System (EI0002), Novex® Sharp Pre-Stained Protein Standard (LC5800), and iBlot® Dry Blotting System (IB21001). Proteins were transferred to a nitrocellulose membrane and blocked with 5% skim milk at 4°C overnight. alpha-Tubulin was detected at ~ 52 kDa using alpha-Tubulin Biotin Conjugated Mouse Monoclonal Antibody (A21371) at 1:3000 dilution in 5% skim milk for 3 hour at room temperature on a rocking platform. Streptavidin - HRP (SA1007) at 1:2000 dilution was used and chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (WP20005).
|Tested species reactivity||Bovine, Human, Mouse|
|Published species reactivity||Not Applicable|
|Host / Isotype||Mouse / IgG1|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:100-1:500|
|Immunocytochemistry (ICC)||1.0 ug/ml|
|Immunofluorescence (IF)||Assay Dependent|
|Immunohistochemistry (Paraffin) (IHC (P))||1:10-1:100|
|Western Blot (WB)||1:1000-1:5000|
* 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.
|Immunocytochemistry (ICC)||See 1 publications below|
Storage and reconstitution: upon receipt, store desiccated at -20°C. To prepare a stock solution, reconstitute in PBS containing 1% BSA and 2mM sodium azide. Reconstituted solutions should remain stable for 6 months when stored at 4°C. For longer storage, aliquot and store at -20°C.
Microtubules of the eukaryotic cytoskeleton perform essential and diverse functions and are composed of a heterodimer of alpha and beta tubulin. The genes encoding these microtubule constituents are part of the tubulin superfamily, which is composed of six distinct families. Genes from the alpha, beta and gamma tubulin families are found in all eukaryotes. The alpha and beta tubulins represent the major components of microtubules, while gamma tubulin plays a critical role in the nucleation of microtubule assembly. There are multiple alpha and beta tubulin genes and they are highly conserved among and between species.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||
A single class II myosin modulates T cell motility and stopping, but not synapse formation.
A21371 was used in immunocytochemistry to analyze modulation of T cell motility and stopping, but not synapse formation by a single class II myosin
|Jacobelli J,Chmura SA,Buxton DB,Davis MM,Krummel MF||Nature immunology (5:531)||2004|