Why is tubulin used in Western blots?
Beta-Tubulin, is usually used as loading control for Western Blot to normalize the levels of protein detected by confirming that protein loading is the same across the gel.
What is the molecular weight of alpha tubulin?
50 kDa
Alpha tubulin molecular weight: 50 kDa.
What is the cellular function of alpha tubulin?
α- and β-tubulins polymerize into microtubules, a major component of the eukaryotic cytoskeleton. Microtubules function in many essential cellular processes, including mitosis.
What is alpha tubulin?
The TUBA1A gene provides instructions for making a protein called alpha-tubulin (α-tubulin). This protein is part of the tubulin family of proteins that form and organize structures called microtubules. Microtubules are rigid, hollow fibers that make up the cell’s structural framework (the cytoskeleton).
Is tubulin a good loading control?
Choosing loading control antibodies Nonetheless, β-actin and β-tubulin offer certain advantages as loading controls: they are highly conserved, display high expression level, and exhibit stability under most experimental conditions.
Is tubulin used as a control?
To date, two housekeeping geneâcoded proteins (i.e., beta-actin and beta-tubulin) are widely used as internal controls in the Western blot analysis.
Is alpha tubulin a good loading control?
When performing a Western Blot, it is crucial to ensure equal loading of protein samples and protein transfer through the use of a loading control antibody. beta-Actin, GAPDH and alpha Tubulin are well known housekeeping proteins that are commonly used as loading controls.
Do nerve cells have gamma-tubulin?
Our studies reveal that, like the situation in nonneuronal cells, gamma-tubulin is restricted to the pericentriolar region of the neuron. Furthermore, serial reconstruction analyses indicate that the minus ends of MTs in both axons and dendrites are free of gamma-tubulin immunoreactivity.
How many kDa is beta actin?
42 kDa
Though beta-actin has a molecular weight of 42 kDa, degradation by apoptosis will produce 38-41 kDa fragments on a Western blot.
What is the function of tubulin protein?
Highly conserved α- and β-tubulin heterodimers assemble into dynamic microtubules and perform multiple important cellular functions such as structural support, pathway for transport and force generation in cell division.
How do I choose a loading control?
Select a loading control that has a different molecular weight to the protein of interest. This ensures that you will be able to distinguish between the bands.
How does the α-tubulin antibody work?
The α-Tubulin Antibody detects endogenous levels of total α-tubulin protein, and does not cross-react with recombinant β-tubulin.
How do you test for α/β tubulin?
Resuspend cells in 200-500 µl of 1X PBS and analyze on flow cytometer. The α/β-Tubulin Antibody detects endogenous levels of both α- and β-tubulin total protein, and does cross-react with both recombinant α- and β-tubulin.
How are pKa C-α and AKT1 antibodies used to detect pKa-C-α?
PKA C-α was detected using the PKA C-α Antibody #4782, and Akt1 was detected using Akt1 (2H10) Monoclonal Antibody #2967. The PKA C-α Antibody confirms silencing of PKA C-α expression, and the Akt1 Antibody is used to control for loading and specificity of PKA C-α siRNA.
How are antibodies purified from microtubules?
Antibodies are purified by protein A and peptide affinity chromatography. Globular tubulin subunits comprise the microtubule building block, with α/β-tubulin heterodimers forming the tubulin subunit common to all eukaryotic cells.