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Beta-Tubulin Loading Control Antibody, Mouse MAb

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Beta-TubulinAntibody Product Information
Antigen:A synthetic peptide corresponding to the N-terminus of the human beta-tubulin
Clone ID:05
Ig Type:Mouse IgG2b
Concentration:
Formulation:0.2 μm filtered solution in PBS with 5% trehalose
Preparation:This antibody was produced from a hybridoma resulting from the fusion of a mouse myeloma with B cells obtained from a mouse immunized with a synthetic peptide corresponding to the N-terminus of the human beta-tubulin. The IgG fraction of the cell culture supernatant was purified by Protein A affinity chromatography.
Beta-TubulinAntibody Usage Guide
Specificity:Human Beta-Tubulin
Application:WB, ICC/IF, IF

WB: 0.5-1 μg/mL

This antibody can be used at 0.5-1 μg/mL with the appropriate secondary reagents to detect Human Beta-Tubulin in WB.

ICC/IF: 5-20 μg/mL

Storage:This antibody can be stored at 2℃-8℃ for one month without detectable loss of activity. Antibody products are stable for twelve months from date of receipt when stored at -20℃ to -70℃. Preservative-Free.
Sodium azide is recommended to avoid contamination (final concentration 0.05%-0.1%). It is toxic to cells and should be disposed of properly. Avoid repeated freeze-thaw cycles.
Images

Immunofluorescence staining of Human Beta-Tubulin in Hela cells. Cells were fixed with 4% PFA, permeabilzed with 1% Triton X-100 in PBS, blocked with 10% serum, and incubated with Mouse anti-Human Beta-Tubulin monoclonal antibody (10 µg/ml) at 4℃ overnight. Then cells were stained with the Alexa Fluor® 594-conjugated (left panel, captured by laser confocal scanning microscope) and Alexa Fluor®488-conjugated (right panel, captured by fluorescence microscope) Goat Anti-mouse IgG secondary antibody, countstained with DAPI (blue). Positive staining was localized to cytoskeleton.

Background

Beta-Tubulin is a subunit of tubulin. Tubulin is one of several members of a small family of globular proteins. It is the major constituent of microtubules. There are two most common members of the tubulin family: alpha-tubulin and beta-tubulin, and together their dimers form microtubules. The dimers of alpha- and beta-tubulin bind to GTP and assemble onto the (+) ends of microtubules while in the GTP-bound state. After the dimer is incorporated into the microtubule, the molecule of GTP bound to the beta -tubulin subunit eventually hydrolyzes into GDP through inter-dimer contacts along the microtubule protofilament. Beta-tubulin faces the plus end of the microtubule while alpha-tubulin faces the minus end. Dimers bound to GTP tend to assemble into microtubules, while dimers bound to GDP tend to fall apart. Loading controls are usually proteins that exhibit high-level, constitutive expression in the cell type or sample you are examining. This ensures constant expression levels. Thus “housekeeping genes” are frequently chosen for use as loading controls. It is also important that the protein chosen as a loading control has a different molecular weight than the protein of interest so that the bands are distinct and expression levels quantifiable. Popular loading control detection antibodies include anti-β-Actin monoclonal or polyclonal antibodies, anti-COX-4, anti-GAPDH, anti-Tubulin and anti-VDAC/Porin antibodies.

References
  • Williams RC Jr. et al., 1999. Anal Biochem. 275(2): 265-7.
  • Nogales E. et al., 1998. Nat Struct Biol. 5(6): 451-8.
  • Dutcher SK. 2001. Curr Opin Cell Biol. 13(1): 49-54.
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