Western blotting is one of the most commonly used antibody assay techniques in cell and molecular biology research since its development over three decades ago, and is considered the gold standard for protein detection and quantification.
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. While there are various options available, researchers often face some challenges in choosing a proper loading control antibody as expression of housekeeping proteins can be influenced by sample type, treatment or other phenomenon. Here are some basic tips to consider when choosing a loading control:
- Detection size: Choose a loading control that does not have a similar molecular weight to that of your protein of interest.
- Strong Expression: Choose a loading control that demonstrates strong expression in your sample.
- Expression Factors: Choose a loading control that shows ubiquitous expression and not affected by an experimental variable.
Please visit our loading controls page for more guidance and to learn more about loading controls.