Anti-alpha-Tubulin Purified

• Catalog Number: orb44543
• Category: Antibodies
• Description: Mouse Monoclonal to alpha tubulin.
• Clonality: Monoclonal
• Clone Number: TU-16
• Tested applications: ELISA, ICC, IHC-P, IP, WB
• Reactivity: Canine, Human, Mouse, Plant, Porcine, Rat
• Isotype: Mouse IgM
• Immunogen: Porcine brain microtubule protein MTP-1.
• Concentration: 1 mg/ml
• Purity: Purified by sequential steps of physicochemical fractionation (differential precipitation and solid-phase chromatography methods).
• Conjugation: Unconjugated
• Target: alpha-Tubulin
• Entrez: 7277
• UniProt ID: Q71U36
• RRID: AB_11012737
• 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-alpha-tubulin antibody
• Note: For research use only
• Application notes: Western blotting: Recommended dilution: 1-2 μg/ml; positive control: HPB-ALL human peripheral blood leukemia cell line, reducing conditions. Immunohistochemistry (paraffin sections): Recommended dilution: 10 μg/ml. Immunoprecipitation: Reducing conditions.
• Expiration Date: 12 months from date of receipt.

Immunoprecipitation of alpha-tubulin from HeLa and RAJI cell lysate by antibody TU-16 and its detection by antibody TU-01. IgM heavy chain (76-92 kDa) and IgM light chain (25-30 kDa) indicated. Mr of alpha tubulin is around 50 kDa. L = lysate; IPr = immunoprecipitate (reducing conditions).

Immunocytochemistry staining of alpha-tubulin in Hep-2 cells using mouse monoclonal antibody TU-16 (diluted 1:400), detected with GAM IgG-Alexa Fluor®488 (diluted 1:200; green).

Immunohistochemistry staining of human heart (paraffin sections) using anti-alpha tubulin (TU-16).

Immunohistochemistry staining (paraffin sections) of alpha-tubulin in human stomach using mouse monoclonal antibody TU-16 (diluted 1:400), detected with GAM IgG-Alexa Fluor®488 (diluted 1:200; green).

Western blotting analysis of human alpha-tubulin using mouse monoclonal antibody TU-16 on lysates of various cell lines and porcine brain 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 54 kDa, nonspecific minor bands above 100 kDa do not interfere with specific signal.

Anti-alpha-Tubulin Purified (TU-16) works in WB application under reducing conditions. 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-16 (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.