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Immunofluorescent analysis of beta-3 Tubulin in SHSY5Y cells (human neuroblast) either left untreated (left panel) or treated with 10uM retinoic acid (right panel) for 72 hours. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature. Cells were blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed without (left panel) or with (right panel) a beta-3 Tubulin monoclonal antibody (Product # MA1-118) at a dilution of 1:100 for at least 1 hour at room temperature, washed with PBS, and incubated with a DyLight 488-conjugated goat anti-mouse IgG secondary antibody (Product # 35502). F-Actin (red) was stained with DyLight-554 Phalloidin (Product # 21834) and nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ToxInsight at 20X magnification.
|Tested species reactivity||Bovine, Guinea pig, Hamster, Human, Mouse, Pig, Rabbit, Rat|
|Host / Isotype||Mouse / IgG2a|
|Immunogen||A synthetic peptide corresponding to amino acids 436-450 from rat neuronal specific beta-3 tubulin.|
|Storage buffer||PBS with 1mg/ml BSA, 30% glycerol|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:50|
|Immunofluorescence (IF)||1:50 - 1:200|
|Immunohistochemistry (IHC)||1:50 - 1:200|
|Immunoprecipitation (IP)||5 µg|
|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.
Western blot analysis of MA1-118 detects an ~50 kDa protein in neuronal-type cells. In bovine, a unknown band at ~32 kDa is also detected. MA1-118 shows specificity to beta-3 Tubulin and is non-reactive to lysates from non-neuronal cell types (e.g. HeLa cell lysate).
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.
beta-4; betaIII-tubulin; CDCBM; CFEOM3A; class III beta-tubulin; neuron-specific class III beta-tubulin; TUBB4; tubulin beta-2B chain; tubulin beta-3 chain; tubulin beta-4 chain; tubulin beta-III; tubulin, beta 3; tubulin, beta 3 class III; tubulin, beta, 3; tubulin, beta, 4
3200002H15Rik; beta-4; BOS_17166; CDCBM; CDCBM1; CFEOM3; CFEOM3A; FEOM3; M(beta)3; M(beta)6; TUBB3; TUBB4