how to choose a proper tag for western blotting - (Oct/01/2014 )

Hi all,

When I did some research about my protein, and found that people always used strep or myc tag for western blotting. However, from my knowledge, his-tag should be the most convinient and popular one.

Do you know why they use these tags? What are differences between those and his-tag in terms of binding specificity like less chance of getting false positive for instance.

Thanks!

Bio

-Biogareth-

His tags are often used for purification of proteins rather than wb. Most common tags used are probably flag and HA.

-bob1-

Thanks Bob1. It is nice to know that. But I am wondering that why tags like you said are often used in wb over histag. What are their advantages?

-Biogareth-

His was designed for column purification and is not particularly antigenic. Flag and ha are more antigenic and strongly hydrophilic.

-bob1-

bob1 on Sun Oct 5 04:49:15 2014 said:

His was designed for column purification and is not particularly antigenic. Flag and ha are more antigenic and strongly hydrophilic.

Hi Bob1,

Thanks for your suggestion. BTW, how about Strep or myc tag?

Best,

Bio

-Biogareth-

Streptavidin is used in conjugation with Biotin, and is a highly specific and sensitive reaction (not an immunologic based reaction though, so it can't be compared to antibody systems). Myc also works well, but I think it has a number of eukaryotic homologues, so you would probably only use it in prokaryotic systems.

-bob1-

Bob1,

Thanks for the information. It is very useful. I am gonna try with Strep first then.

Bio

-Biogareth-