Mouse Monoclonal to Alpha Tubulin Antibody from Abcam

Abcam’s mouse monoclonal to alpha Tubulin antibody is an excellent protein loading control antibody.

  • Product Name - Anti-alpha Tubulin antibody [DM1A]—Loading Control
  • Description - Mouse monoclonal [DM1A] to alpha Tubulin—Loading Control
  • Host Species - Mouse
  • Tested Applications - DM1A is best suited for ICC/IF, Flow Cyt, IHC-Fr, IP, Electron Microscopy, IHC-P, and WB.
  • Species Reactivity - DM1A reacts with rat, mouse, guinea pig, chicken, cow, hamster, humans, dog, Xenopus laevis, pig, African green monkey, and gerbil.
  • Immunogen - Full-length native protein (purified) corresponding to chicken alpha Tubulin.
  • Epitope - aa 426-450

Specificity

DM1A makes the 10 nm filaments to disintegrate into large lateral aggregates that get collected in tight juxtanuclear caps or the cell periphery. It neither suppresses polymerization or depolymerization of platelet tubulin in vitro nor blocks microtubule assembly. DM1A blocks, by 70%–80%, the potential of tubulin dimers (including GppNHp bound) to stimulate a stable inhibition of adenylyl cyclase.

Positive Control

  • Flow cytometry: methanol fixed/Tween permeabilized HeLa cells
  • WB: HEK293, HeLa, Caco-2, HepG2, PC12, NIH3T3
  • IHC-P: Rat colon, human colon
  • ICC/IF: SV40LT-SMC, NIH3T3, Caco-2

Applications

Abcam’s Abpromise guarantee covers the usage of ab7291 in the tested applications listed in the table below. Note: Recommended starting dilutions have been mentioned; the end user should determine the ideal concentrations/dilutions.

Application Abreviews Notes
Flow Cyt 1 µg should be used for 106 cells.
The ab170190 Mouse monoclonal IgG1 is ideal for use as an isotype control with this antibody.
ICC/IF ☆☆☆☆☆ A concentration of 0.5–1 µg/mL should be used.
IP Should be used at an assay dependent concentration.
IHC-Fr ☆☆☆☆ Should be used at an assay dependent concentration.
IHC-P ☆☆☆☆☆ A concentration of 5 µg/mL should be used. Heat-mediated antigen retrieval should be performed with citrate buffer pH 6 prior to starting with IHC staining protocol.
Electron Microscopy Should be used at an assay dependent concentration.
WB ☆☆☆☆☆ 1/5000–1/10000. A band of approximately 50 kDa (predicted molecular weight: 50 kDa) is detected.

Recommendations are provided to dilute ab7291 to 1:10000 and incubate it overnight at 4 °C. It functions under both reducing and non-reducing conditions. It is recommended to use 3% BSA as the blocking agent; blocking with milk may lead to a reduction in signal intensity.

Target

Function

Tubulin is the important component of microtubules that binds two moles of GTP—one at a non-exchangeable site on the alpha chain and the other at an exchangeable site on the beta chain.

Sequence Similarities

It belongs to the tubulin family.

Post-Translational Modifications

Certain glutamate residues located at the C-terminus are polyglutamylated, where the modification takes place particularly on glutamate residues and leads to polyglutamate chains on the gamma-carboxyl group. Moreover, the glutamate residues are monoglycylated and not polyglycylated because of the lack of functional TTLL10 in humans.

Monoglycylation is primarily restricted to tubulin integrated into axonemes (flagella and cilia), while glutamylation is common in centrioles, neuronal cells, the mitotic spindle, and axonemes. There is a possibility of both modifications coexisting on the same protein on adjoining residues. A drop in the glycylation levels leads to an increase in polyglutamylation, and vice versa.

Although the exact role of such modifications is still ambiguous, they govern the dynamics and assembly of axonemal microtubules.

Acetylation of alpha chains at Lys-40 leads to stabilization of microtubules and has an impact on the processivity and affinity of microtubule motors. This modification plays a role in a number of cellular functions such as cell motility, cell cycle progression, or cell differentiation to intracellular signaling and trafficking.

Cellular Localization

Cytoplasm > cytoskeleton