Anti-alpha-Tubulin Purified

• Catalog Number: orb44534
• Category: Antibodies
• Description: Mouse Monoclonal to alpha tubulin.
• Target: alpha-Tubulin
• Clonality: Monoclonal
• Isotype: Mouse IgM
• Conjugation: Unconjugated
• Reactivity: Human, Mouse, Plant, Porcine
• Concentration: 1 mg/ml
• Buffer/Preservatives: Tris buffered saline (TBS), pH 8.0, 15 mM sodium azide
• Purity: Purified by sequential steps of physicochemical fractionation (differential precipitation and solid-phase chromatography methods).
• Immunogen: microtubule proteins from porcine brain
• UniProt ID: Q71U36
• RRID: AB_10993590
• Tested applications: WB
• Application notes: Western blotting: Recommended dilution: 1-2 μg/ml.
• Antibody Type: Primary Antibody
• Clone Number: TU-02
• 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.
• Alternative names: Anti-alpha-tubulin antibody
• Note: For research use only
• Entrez: 7277

Western blotting analysis of human alpha-tubulin using mouse monoclonal antibody TU-02 on lysates (50 mM TRIS-Cl pH 6.8, 4M UREA, 4% SDS) of various cell lines under reducing and non-reducing conditions. Nitrocellulose membrane was probed with 2 µg/ml of mouse anti-alpha-tubulin monoclonal antibody followed by IRDye800-conjugated anti-mouse secondary antibody. A specific band was detected for alpha-tubulin at approximately 55 kDa.

Anti-alpha-Tubulin Purified (TU-02) 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, Jurkat, and Raji cell lines mixed and heated (100°C, 5 min) with reducing (2-mercaptoethanol) or non-reducing SDS-loading buffer. Samples were resolved using 15% Tris-glycine SDS gel electrophoresis. Nitrocellulose membrane blot was probed simultaneously with mouse IgM monoclonal antibody TU-02 (1 µg/ml) and anti-GAPDH mouse IgG1 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. Anti-alpha-Tubulin Purified (TU-02) is not suitable for use in non-reducing conditions on RIPA cell extracts.