Anti-Alpha-Tubulin [F2C]

• Catalog Number: orb411570
• Category: Antibodies
• Description: Human monoclonal antibody to Alpha-Tubulin
• Species/Host: Human
• Clonality: Monoclonal
• Clone Number: F2C
• Tested applications: ELISA, WB
• Reactivity: Human
• Isotype: Human IgG
• Immunogen: Human MBP (microtubule-binding protein).
• Concentration: 1 mg/ml
• Purity: Purified
• Conjugation: Unconjugated
• Target: Alpha-Tubulin
• UniProt ID: Q71U36
• 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: PBS with 0.02% Proclin 300.
• Alternative names: TUBA
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Flow-cytometry using the anti-Alpha Tubulin F2C HepG2 (A), HeLa (B) and Hek293 (C) cells were stained with unimmunized rabbit IgG antibody (black line) or the rabbit-chimeric version of F2C (orb411570, blue line) at a concentration of 10 µg/ml for 30 mins at RT. After washing, bound antibody was detected using anti-rabbit IgG JK (FITC-conjugate) antibody (129936) at 2 µg/ml and cells analyzed on a FACSCanto flow-cytometer.

Immunofluoresence staining of fixed HeLa cells with anti-Alpha-tubulin antibody F2C. Immunofluorescence analysis of paraformaldehyde fixed HeLa cells, permeabilized with 0.15% Triton stained with the chimeric rabbit IgG version of F2C (orb411570) at 10 µg/ml for 1h followed by Alexa Fluor® 488 secondary antibody (1 µg/ml), showing microtubule staining. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom orb411570, DAPI, merged channels and a negative control. The negative control was stained with unimmunized rabbit IgG followed by Alexa Fluor® 488 secondary antibody.

Immunohistochemical staining of human cerebral cortex tissue using anti-Alpha Tubulin antibody F2C Anti-Alpha Tubulin staining of paraffin embedded human cerebral cortex tissue using the rabbit-chimeric version of F2C (orb411570). Antigen retreival was acheived by microwaving in citrate buffer (pH6), followed by blocking with protein block serum-free buffer. Primary antibody incubation with orb411570 was carried out at 4 µg/ml for 30 minutes. Samples were then incubated with an anti-rabbit IgG HRP secondary antibody for 20 mins followed by DAB (3, 3′-diaminobenzidine), and counter-staining with haemotoxylin. Staining of axons and a few neuronal cell bodies may be observed. Recommended concentration, 2-4 µg/ml.

Western Blot using anti-Alpha-Tubulin antibody F2C HeLa (A), HEK293 (B), and HepG2 (C) cell lysate samples (35 µg protein in RIPA buffer) were resolved on a 10% SDS PAGE gel and blots probed with the chimeric rabbit version of F2C (orb411570) at 0.01 µg/ml before detection using an anti-rabbit secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence. The expected band size for Alpha-Tubulin is ~55kDa. orb411570 successfully detected human Alpha-Tubulin in HeLa, HEK293, and HepG2 cell lysate samples.