Immunofluorescence analysis of beta Tubulin was performed using 70% confluent log phase SH-SY5Y 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 Tubulin (38F4) Mouse Monoclonal Antibody (480011) at 1:250 dilution in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A28175) 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 (S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # 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.
|Tested species reactivity||Bovine, Human, Mouse, Rat|
|Published species reactivity||Mouse, Not Applicable|
|Host / Isotype||Mouse / IgG|
|Immunogen||Synthetic peptide corresponding to beta III tubulin|
|Storage buffer||0.01M HEPES, pH 7.5, with 0.15M NaCl, 100µg/ml BSA, 50% glycerol|
|Storage Conditions||-20°C or -80°C if preferred|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunohistochemistry (Paraffin) (IHC (P))||Assay Dependent|
|Western Blot (WB)||1:5000 - 1:10000|
* 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.
Beta tubulins are one of two core protein families (alpha and beta tubulins) that heterodimerize and assemble to form microtubules. Beta-III tubulin is primarily expressed in neurons and may be involved in neurogenesis, axon guidance and maintenance. Mutations in this gene are the cause of congenital fibrosis of the extraocular muscles type 3. Beta-III tubulin was also detected in Sertoli cells of the testis and transiently in non-neuronal embryonic tissues. Glutamate residues at the C-terminus of beta III tubulin can be glutamylated. The precise function of such modifications is unclear. Tubulin is phosphorylated on Ser-172 by CDK1 during cell cycle progression. Ser-172 phosphorylation inhibits tubulin incorporation into microtubules.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
FHL1 reduces dystrophy in transgenic mice overexpressing FSHD muscular dystrophy region gene 1 (FRG1).
480011 was used in western blot to assess how FRG1 expression contributes to myoblast differentiation defects
|Feeney SJ,McGrath MJ,Sriratana A,Gehrig SM,Lynch GS,D'Arcy CE,Price JT,McLean CA,Tupler R,Mitchell CA||PloS one (10:null)||2015|
Major components of energy drinks (caffeine, taurine, and guarana) exert cytotoxic effects on human neuronal SH-SY5Y cells by decreasing reactive oxygen species production.
480011 was used in immunocytochemistry to assess if major components of energy drinks exert cytotoxic effects on human neuronal SH-SY5Y cells
|Zeidán-Chuliá F,Gelain DP,Kolling EA,Rybarczyk-Filho JL,Ambrosi P,Terra SR,Pires AS,da Rocha JB,Behr GA,Moreira JC||Oxidative medicine and cellular longevity (2013:null)||2013|