Immunofluorescence analysis of Beta-3 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-3 Tubulin (TU-20) Mouse Monoclonal Antibody (MA119187) at 2ug/ml 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||Many|
|Published species reactivity||Human|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Peptide (C) 441-448 coupled to maleimide-activated keyhole limpet hemocyanin via cysteine added to the N-terminus of the neuron-specific peptide|
|Purification||Ammonium sulfate precipitation, Caprylic acid precipitation|
|Storage buffer||PBS, pH 7.4|
|Contains||15mM sodium azide|
|Storage Conditions||4° C, do not freeze|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3 - 5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||10 µg/ml|
|Western Blot (WB)||1-2 µg/ml, 90 min|
* 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.
This antibody recognizes the C-terminal peptide sequence ESESQGPK (aa 441-448) of neuron-specific human beta III-tubulin.
For immunohistochemistry applications, this product requires protein pretreatment (0.1% trypsin) in 0.1 M HCl; incubate for 30 minutes at room temperature.
Antibodies to this protein (and modification) were previously sold as part of a Thermo Scientific Cellomics High Content Screening Kit. This replacement antibody is now recommended for researchers who need an antibody for high content cell based assays. It has been thoroughly tested and validated for cellular immunofluorescence (IF) applications. Further optimization including the selection of the most appropriate fluorescent Dylight conjugated secondary antibody may have to be performed for your high content assay.
The betaIII-tubulin isoform is present dominantly in cells of neuronal origin and it is one of the earliest markers of neuronal differentiation. It is regarded as a specific probe for the cells of neuronal origin as well as for the tumours originating from these cells. The betaIII-tubulin is most abundant in cells of neuronal origin, but was also detected in Sertoli cells of the testis and transiently in non-neuronal embryonic tissues.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Progerin impairs chromosome maintenance by depleting CENP-F from metaphase kinetochores in Hutchinson-Gilford progeria fibroblasts.
MA1-19187 was used in western blot test if progerin elicits spatiotemporal deviations in mitotic processes in Hutchinson-Gilford progeria syndrome fibroblasts
|Eisch V,Lu X,Gabriel D,Djabali K||Oncotarget (7:24700)||2016|
Novel crystalloid oligodendrogliopathy in hereditary spastic paraplegia.
MA1-19187 was used in immunohistochemistry to study crystalloid oligodendrogliopathy in hereditary spastic paraplegia
|Woehrer A,Laszlo L,Finsterer J,Stöllberger C,Furtner J,Rinner W,Molnar K,Budka H,Kovacs GG||Acta neuropathologica (124:583)||2012|