TUBB3 antibody (AA 443-450)

Catalog No. ABIN1742553

Product Details anti-TUBB3 Antibody

Target: TUBB3 (Tubulin, beta 3 (TUBB3))

Binding Specificity: AA 443-450

Reactivity: Human, Mouse, Rat

Host: Rabbit

Clonality: Polyclonal

Conjugate: This TUBB3 antibody is un-conjugated

Application: Western Blotting (WB), Immunohistochemistry (IHC), Immunoprecipitation (IP), Immunocytochemistry (ICC)

Specificity: Specific for beta3-tubulin.

Purification: antiserum

Immunogen: Synthetic peptide (aa 443-450 of mouse beta3-tubulin) coupled to key-hole limpet hemocyanin via an added N-terminal cysteine.

Application Details

Application Notes: WB: 1 : 1000 up to 1 : 10000 (AP staining)
ICC: 1 : 1000 up to 1 : 5000
IHC: 1 : 200 up to 1 : 500

Restrictions: For Research Use only

Independent Validation (WB)

Validation #029822 (Western Blotting)

by: Kinexus Bioinformatics Corporation

No.: #029822

Date: 10/28/2014

Lot Number: 302302/3

Method validated: Western Blotting

Positive Control: Mouse brain

Negative Control: Mouse heart, HeLa cells, A431 cells

Notes: A band was observed in the positive controls at the expected size (~50 kDa), which is lower in the negative controls.

Validation Images

Figure 1: Western Blot for Tubulin, Beta, 3 (TUBB3). Grey arrowhead indicates the expected molecular weight of ~50 kDa.

Full Methods

Primary Antibody:

• Antigen: Tubulin, Beta, 3 (TUBB3) (AA 443-450)
• Catalog number: ABIN1742553
• Lot number: 302302/3
• Dilution: 1:1000

Secondary Antibody:

• Antibody: Donkey anti-Rabbit IgG Antibody (HRP)
• Lot number: F0613
• Dilution: 1:10,000

Full Protocol:

• 1. Cell/tissue total protein lysates were boiled in 1X SDS Sample Buffer containing 1% SDS and 1.25% β-mercaptoethanol at 95°C for 5 minutes prior to loading.
• 2. 15 μg of boiled lysate were loaded and resolved on a 12% SDS-polyacrylamide gel.
• 3. The Precision Plus Protein™ All Blue Prestained Standards from BioRad (161-0373) were used as molecular mass markers.
• 4. Proteins were transferred onto nitrocellulose membrane by tank transfer and protein transfer was confirmed with Ponceau S staining.
• 5. The immunoblot membrane was blocked in 2.5% skim milk and 1.5% BSA solution in TTBS at room temperature for 60 minutes.
• 6. The membrane was washed in TTBS twice for 5 minutes each.
• 7. The membrane was immersed with the protein side up in the antibody solution in TBS and incubated overnight at 4°C with gentle agitation.
• 8. The membrane was rinsed twice with TTBS.
• 9. The membrane was washed in TTBS twice for 5 minutes each.
• 10. The membrane was washed in TTBS once for 15 minutes.
• 11. The membrane was incubated in the HRP-conjugated secondary antibody solution in TBS for 60 minutes at room temperature with gentle agitation.
• 12. The membrane was rinsed twice with TTBS.
• 13. The membrane was washed in TTBS twice for 5 minutes each.
• 14. The membrane was washed in TTBS once for 15 minutes.
• 15. Signals were detected by chemiluminescence (ECL). The blot was scanned for 320 seconds.
• 16. The membrane was rinsed three times with TTBS.
• 17. Repeated Steps 4-15 with the loading control antibody and its matching secondary antibody. The blot was scanned for 160 seconds.

Experimental Notes: - No experimental challenges noted.

Handling

Format: Liquid

Buffer: PBS

Handling Advice: Crude antisera are more robust than monoclonals. With anti-microbials added, they may be stored at 4 °C.
Serum does not contain active proteases, in fact, serum itself contains a powerful cocktail of protease inhibitors. Frozen storage (-20 °C),however, is preferable.

Storage: 4 °C/-20 °C

Storage Comment: Unlabeled antibodies are stable in this form without loss of quality at ambient temperatures for several weeks or even months. They can be stored at 4 °C for several years.

References for anti-TUBB3 antibody (ABIN1742553)

Kaczor, Rakus, Mozrzymas: "Neuron-astrocyte interaction enhance GABAergic synaptic transmission in a manner dependent on key metabolic enzymes." in: Frontiers in cellular neuroscience, Vol. 9, pp. 120, (2015) (PubMed).

Target Details for TUBB3

Target: TUBB3 (Tubulin, beta 3 (TUBB3))

Alternative Name: beta3-Tubulin (TUBB3) (TUBB3 Products)

Synonyms: CDCBM antibody, CFEOM3A antibody, TUBB4 antibody, beta-4 antibody, TUBB3 antibody, 3200002H15Rik antibody, M(beta)3 antibody, M(beta)6 antibody, tubb3 antibody, tubulin beta 3 class III antibody, tubulin, beta 3 class III antibody, tubulin beta 3 class III L homeolog antibody, beta-tubulin 3 antibody, tubulin beta-3 chain antibody, beta tubulin 3 antibody, TUBB3 antibody, Tubb3 antibody, tubb3 antibody, tubb3.L antibody, LOC100581245 antibody, tub3 antibody

Background: Synonyms: TuJ1

Pathways: Microtubule Dynamics, M Phase