This antibody recognizes the C-terminal peptide sequence ESESQGPK (aa 441-448) of Human class III β-tubulin specific for neurones. The antibody is a highly specific marker for neuronal tissue. TU-20 is very useful for the detection of microtubule structures on fixed cells. This Monoclonal antibody TU-20 is widely cross-reactive among species (recognized epitope conserved within all species).
Purification
Ammonium Sulphate and Caprylic Acid Precipitation
Immunogen
Peptide (C) 441-448 coupled to Maleimide-activated Keyhole Limpet Hemocyanin via Cysteine added to the N-terminus of the neuron-specific peptide.
TUBB3
Reactivity: Human
WB, ELISA, IHC, IF, IP
Host: Mouse
Monoclonal
unconjugated
Application Notes
Flow cytometry. ELISA. Western Blotting: 1-2 μg/mL, 90 min. Positive Control: Porcine brain lysate. Negative Control: HPB-ALL peripheral blood leukemia cell line. Sample Preparation: Mix lysate with reducing Laemmli SDS-PAGE sample buffer. Application note: Reducing conditions. Immunohistochemistry on Frozen Sections. Immunohistochemistry on Paraffin Sections: 10 μg/mLStaining technique: Standard ABC technique (DAB+). Pretreatment: 0.1 % pepsin (trypsin) in 0.1 M HCl, incubation 30 min in RT or hightemperature citrate buffer antigen retrieval. Positive tissue: Neuronal tissue. Immunocytochemistry: FITC conjugate: 3 μg/mL. Positive Material: Neuro2a mouse neuroblastoma cell line. Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.
Restrictions
For Research Use only
Concentration
1.0 mg/mL
Buffer
PBS, pH ~7.4, 15 mM Sodium Azide
Preservative
Sodium azide
Precaution of Use
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice
Avoid repeated freezing and thawing.
Storage
4 °C/-20 °C
Storage Comment
Store undiluted at 2-8 °C for one month or (in aliquots) at -20 °C for longer.
Katsetos, Draber, Kavallaris: "Targeting βIII-tubulin in glioblastoma multiforme: from cell biology and histopathology to cancer therapeutics." in: Anti-cancer agents in medicinal chemistry, Vol. 11, Issue 8, pp. 719-28, (2012) (PubMed).
Theodorou, Dalembert, Heffelfinger, White, Weissman, Corcoran, Snyder: "A high throughput embryonic stem cell screen identifies Oct-2 as a bifunctional regulator of neuronal differentiation." in: Genes & development, Vol. 23, Issue 5, pp. 575-88, (2009) (PubMed).
Dráberová, Del Valle, Gordon, Marková, Smejkalová, Bertrand, de Chadarévian, Agamanolis, Legido, Khalili, Dráber, Katsetos: "Class III beta-tubulin is constitutively coexpressed with glial fibrillary acidic protein and nestin in midgestational human fetal astrocytes: implications for phenotypic identity." in: Journal of neuropathology and experimental neurology, Vol. 67, Issue 4, pp. 341-54, (2008) (PubMed).
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Background
The beta-III Tubulin isotype is present dominantly in cells of neuronal origin and it is one of the earliest marker of neuronal differentiation. Class III beta-tubulin, is regarded as a specific probe for the cells of neuronal origin as well as for the tumours originating from these cells. The neuron-associated class III beta-tubulin isotype 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.Synonyms: Tubulin beta-3 chain, Tubulin beta-4, Tubulin beta-III