Anti-beta-Tubulin Purified

• Catalog Number: orb44535
• Category: Antibodies
• Description: Mouse Monoclonal to beta tubulin.
• Clonality: Monoclonal
• Clone Number: TU-06
• Tested applications: ICC, IHC-P, WB
• Reactivity: Fish, Gallus, Human, Mouse, Other, Paramecium, Plant, Porcine, Rat
• Isotype: Mouse IgM
• Immunogen: Beta-subunits of porcine brain tubulin.
• Concentration: 1 mg/ml
• Purity: Purified by sequential steps of physicochemical fractionation (differential precipitation and solid-phase chromatography methods).
• Conjugation: Unconjugated
• Target: beta-Tubulin
• Entrez: 81027
• UniProt ID: Q9H4B7
• RRID: AB_10990495
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: Tris buffered saline (TBS), pH 8.0, 15 mM sodium azide
• Alternative names: Anti-beta-tubulin antibody
• Note: For research use only
• Application notes: Immunocytochemistry: Recommended dilution: 2 μg/ml, fixed and permeabilized cells; positive control: 3T3 mouse embryonal fibroblast cell line. Immunohistochemistry (paraffin sections): Recommended dilution: 5 μg/ml, positive tissue: heart. Western blotting: Recommended dilution: 1-2 μg/ml.
• Expiration Date: 12 months from date of receipt.

Immunocytochemistry staining of 3T3 mouse embryonal fibroblast cell line using anti-beta-tubulin (TU-06) (detection by Goat anti-mouse IgM Cy®5). Nucleus is stained with DAPI (blue).

Immunohistochemistry staining of human heart (paraffin sections) using anti-beta tubulin (TU-06).

Western blotting analysis of human beta-tubulin using mouse monoclonal antibody TU-06 on lysates (50 mM TRIS-Cl pH 6.8, 4M UREA, 4% SDS) of various cell lines under non-reducing and reducing conditions. Nitrocellulose membrane was probed with 2 µg/ml of mouse anti-beta-tubulin monoclonal antibody followed by IRDye800-conjugated anti-mouse secondary antibody. A specific band was detected for beta-tubulin at approximately 54 kDa.

Anti-beta-Tubulin Purified (TU-06) works in WB application under reducing conditions on RIPA cell extracts. Western blotting analysis was performed on whole cell extracts (RIPA lysis buffer) of HeLa, HEK 293, ESS-1 and Jurkat cell lines mixed and heated (100°C, 5 min) with reducing (2-mercaptoethanol) or non-reducing SDS-loading buffer. Samples were resolved using 12% Tris-glycine SDS gel electrophoresis. Nitrocellulose membrane blot was probed simultaneously with mouse IgM monoclonal antibody TU-06 (1 µg/ml) and mouse IgG1 anti-GAPDH monoclonal antibody FF26A (1 µg/ml) used as the loading control. Subclass-specific secondary antibodies IRDye 680RD Goat-anti-Mouse IgM (red) and IRDye 800CW Goat-anti-Mouse IgG (green) were used for multiplex fluorescent Western blot detection. Alpha-tubulin was detected at ~50 kDa in all tested cell lines under reducing, but not under non- reducing conditions. Using RIPA lysis buffer in combination with non-reducing conditions is not suitable for Anti-beta-Tubulin Purified (TU-06).