Anti-alpha-Tubulin Purified

• Catalog Number: orb44529
• Category: Antibodies
• Description: Mouse Monoclonal to alpha tubulin.
• Clonality: Monoclonal
• Clone Number: TU-01
• Tested applications: FC, ICC, IHC-P, IP, WB
• Reactivity: Aves, Human, Invertebrate, Mouse, Paramecium, Plant, Porcine, Yeast
• Isotype: Mouse IgG1
• Immunogen: Fraction of tubulin purified from porcine brain by two cycles of polymerization - depolymerization.
• Concentration: 1 mg/ml
• Purity: Purified by protein-A affinity chromatography.
• Conjugation: Unconjugated
• Target: alpha-Tubulin
• Entrez: 7277
• UniProt ID: Q71U36
• RRID: AB_11021844
• 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: Phosphate buffered saline (PBS), pH 7.4, 15 mM sodium azide
• Alternative names: Anti-alpha-tubulin antibody
• Note: For research use only
• Application notes: Flow cytometry: Recommended dilution: 1-4 μg/ml, intracellular staining. Western blotting: Recommended dilution: 1-2 μg/ml, 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); IPn = immunoprecipitate (non-reducing conditions).

Separation of HeLa cells stained using anti-alpha-Tubulin (TU-01) purified antibody (concentration in sample 3 µg/ml, GAM APC, red-filled) from HeLa cells unstained by primary antibody (GAM APC, black-dashed) in flow cytometry analysis (intracellular staining).

Immunocytochemistry staining of 3T3 mouse embryonal fibroblast cell line using anti-alpha-tubulin (TU-01; green) and anti-Vimentin (VI-01; cat. no. orb44570; red). Nucleus is stained with DAPI (blue).

Immunocytochemistry staining of HeLa human cervix carcinoma cell line using anti-alpha-tubulin (TU-01; red). Nucleus is stained with DAPI (blue).

Immunocytochemistry staining of 3T3 mouse embryonal fibroblast cell line using anti-alpha-tubulin (TU-01; green). Nucleus is stained with DAPI (blue).

Immunohistochemistry staining of human heart (paraffin sections) using anti-alpha-tubulin (TU-01).

Western blotting analysis of human alpha-tubulin using mouse monoclonal antibody TU-01 on lysates 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 54 kDa.

Use of anti-alpha-tubulin antibody TU-01 as a loading control (A) in an Western blotting experiment revealing the staining pattern of various cell lysates by a newly developed monoclonal antibody (B).

Anti-alpha-Tubulin Purified (TU-01) works in WB application under reducing conditions. Western blotting analysis was performed on whole cell extracts (RIPA lysis buffer) of JAR, JEG3, HTr-8/SVneo, and HeLa cell lines mixed and heated (100°C, 5 min) with reducing (2-mercaptoethanol) or non-reducing SDS-loading buffer. Samples were resolved using 10% Tris-glycine SDS gel electrophoresis. Nitrocellulose membrane blot was probed simultaneously with mouse IgG1 monoclonal antibody TU-01 (1 µg/ml) and mouse IgM monoclonal antibody VI-01 detecting vimentin. Subclass-specific secondary antibodies IRDye 800CW Goat-anti-Mouse IgG (green) and IRDye 680RD Goat-anti-Mouse IgM (red) were used for multiplex fluorescent Western blot detection. Alpha-tubulin was detected at ~50 kDa and vimentin at ~55 kDa.