I am interested in opinions / suggestions on reagents to detect affinity tags on a Western blot. In particular, I am looking to detect 6 x His and Strep tags and in the future GST and MBP tags.
Do people still use chromogenic detection or is it all in chemiluminescence?
For His tag/GST tag, ECL detection (chemiluminescence) worked fine for me always! One has to really really cautious about choosing anti-His antibody (monoclonal preferably) and validating the data since there is a great risk on detecting His rich E.coli protein (slyD chaperone for example). Good luck!
I guess I'm really asking more about the primary detection - the development method is a secondary concern.
Are there universally accepted anti-His, anti-Strep, anti-GST, etc. reagents?
Usually you would use peroxidase coupled to the respective affinity ligand. Such as Nickel-HRP, Glutathione-HRP, ... Then, pick a peroxidase substrate for your preferred detection optics, like TMB for chromogen, ECL for luminescence, and they have fluorescent ones too. All that should be commercially available.
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