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HA tag not visible in Arabidopsis samples

HA tag arabidopsis

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#1 CNJacott



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Posted 20 March 2015 - 08:58 AM



I have recently been carrying out western blots on Arabidopsis thaliana leaf samples.

The gene of interest is overexpressed using a 35S promoter and is tagged to an HA epitope (with no spacer region between).

I am unable to detect any HA protein in my samples. To confirm the function of the Anti-HA primary and secondary antibodies, I used a positive control of a purified HA protein. This did show up on my Western Blots. However this protein was not a protein produced in planta. The antibodies used have apparently worked previously in in tobacco.

Also, I am able to detect the overexpression of the gene of interest using RT-qPCR so it should be present in my samples.

Has anybody got any suggestions as to why I am unable to detect HA in my sample?

Thank you for your help



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