TUBB3 antibody (AA 443-450)

Catalog No. ABIN1742553

This anti-TUBB3 antibody is a Rabbit Polyclonal antibody detecting TUBB3 in WB, IHC, IP and ICC. Suitable for Human, Mouse and Rat. Independently validated for use in Western Blotting. This Primary Antibody has been cited in 1 publication.

Quick Overview for TUBB3 antibody (AA 443-450) (ABIN1742553)

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

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)

Product Details anti-TUBB3 Antibody

Binding Specificity: AA 443-450

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)

Background: Synonyms: TuJ1

Pathways: Microtubule Dynamics, M Phase