What is the meaning of the two bands of HIf 1 alpha in western blot? Why sometimes it appears like two bands? thanks!
Hello María Isabel Hernández Cortés
HIF-1 alpha can be a challenging target to work with in Western Blot. This is because HIF-1 alpha is only stabilized at O2 concentrations below 5%. Under normoxic conditions, HIF-1 alpha has a short half-life and may be degraded within 5-8 minutes in both nuclear and cytoplasmic compartments. Therefore, proper sample preparation is critical for Western Blot to be successful.
Sometimes the observed band size of HIF-1 alpha may not be as predicted in Western Blot due to different forms of HIF-1 alpha which include:
1)40-80 kDa which indicates degraded HIF-1 alpha,
2)110-130 kDa which indicates post-translationally modified HIF-1 alpha, and
3)~200 kDa which indicates heterodimer with HIF-1 beta
Best.
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