TUBB3 / TUBB4 (Loading Control) Antikörper

Antigen: TUBB3 / TUBB4 (Loading Control)

Klonalität: Monoklonal (TU-20)

Wirt: Maus

Applikation: Immunhistochemie (Gefrierschnitte) (IHC (fro)), Enzyme Immunoassay (EIA), Immunhistochemie (Paraffinschnitte) (IHC (p)), Western Blot (WB), Durchflusszytometrie (FACS), Immunfluoreszenz (IF)

Produktnummer: ABIN110980

Produktbeschreibung

Produktmerkmale: Synonyms: Tubulin beta-3 chain, Tubulin beta-III, Tubulin beta-4

Swiss-Prot: Q13509

Immunogen: Synthetic C-terminal peptide

Isotyp: IgG1  (Passende Sekundärantikörper)

Klon: TU-20

Beschreibung: Tubulin is the major building block of microtubules. This intracellular cylindricalfilamentous structure is present in almost all eukaryotic cells. Microtubules function asstructural and mobile elements in mitosis, intracellular transport, flagellar movement, andthe cytoskeleton. Except in the simplest eukaryotes, tubulin exists in all cells as a mixtureof similar, but not identical, sets of alpha and beta tubulin polypeptides. Within either setof polypeptides, individual subunits diverge from each other (both within and acrossspecies) at less than 10% of the amino acid positions. The most extreme diversity islocalized to the 15 residues of the carboxy terminal. For beta tubulin five evolutionarilyconserved isotype clones have been identified. These are almost totally conserved in thesubunits utilized in the same cell types of different species, with the exception of thehematopoietic beta tubulin which is the most highly divergent in sequence and is notconserved between species. Research has been centered around the hypothesis that thesebeta tubulin isotypes contribute to unique functional properties. It has been reported thatthe different isotypes of tubulin differ from each other in their ability to polymerize intomicrotubules. The monoclonal antibody from hybridoma SDL. 3D10 can stimulatemicrotubule assembly when reconstituted with tubulin, tau or MAP2.

Spezifität: The antibody is a highly specific marker for neuronal tissue. It reacts with the class III betaisoform of Tubulin, peptide ESESQGPK specific for neurons. TU-20 is very useful for the detection of microtubule structures on fixed cells. Species: Human and Rat. Others not tested.

Anwendungen

Protokoll: Protocol with frozen, ice-cold acetone-fixed sections: The whole procedure is performed at room temperature1. Wash in PBS2. Block endogenous peroxidase3. Wash in PBS4. Block with 10% normal goat serum in PBS for 30min. in a humid chamber5. Incubate with primary antibody (dilution see datasheet) for 1h in a humid chamber6. Wash in PBS7. Incubate with secondary antibody (peroxidase-conjugated goat anti mouse IgG+IgM(H+L) minimal-cross reaction to human) for 1h in a humid chamber8. Wash in PBS9. Incubate with AEC substrate (3-amino-9-ethylcarbazol) for 12min. 10. Wash in PBS11. Counterstain with Mayer’s hemalumProtocol with formalin-fixed, paraffin-embedded sections: The whole procedure is performed at room temperature1. Deparaffinize and rehydrate tissue section2. Incubate the tissue section with proteinase K for 7min. 3. Wash in distilled water4. Block endogenous peroxidase5. Wash in PBS6. Block with 10% normal goat serum in PBS for 30min. in a humid chamber7. Incubate with primary antibody (dilution see datasheet) for 1h in a humid chamber8. Wash in PBS9. Incubate with secondary antibody (peroxidase-conjugated goat anti mouse IgG+IgM(H+L) minimal-cross reaction to human) for 1h in a humid chamber10. Wash in PBS11. Incubate with AEC substrate (3-amino-9-ethylcarbazol) for 12min. 12. Wash in PBS13. Counterstain with Mayer’s hemalum

Applikationshinweise: Immunohistochemistry on Frozen Sections: 5-10 µg/ml (1/40-1/80). Immunohistochemistry on Paraffin Sections: 20 µg/ml (1/20). Proteinase K treatment forantigen retrieval is recommended. Recommended Positive Control: Human cortex. Has been described to work in ELISA and Western blot. Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.

Konzentration: 0.4 mg/ml

Buffer: Stock solution contains PBS pH 7.2 with 0.09% Sodium Azide aspreservative and 5 mg/ml BSA as stabilizer.Reconstitution: Restore in 0.5 ml distilled water.

Lagerung: Original vial: 1 year at 2-8°C. Aliquots of stock solution can be kept frozen at -20°C, do not freeze working dilutions! Avoid repeated freezing and thawing. Shelf life: one year at -20°C from despatch.

Beschränkungen: Nur für Forschungszwecke einsetzbar