I bought the HisProbe-HRP system from Thermo. First experiments showed very very nice bands and it is much more sensitive than anti-6xHis-Antibodys. But it is recommended using BSA as a blocking reagent. Has someone experience with the HisProbe system using (5%) milk powder instead? Although it's quite "cheap" (I know, bad word), I only want to give it a shot if someone gets data with milk. Thank you in advance!
What is in the hisProbe? Is this a Nickel conjugated secondary antibody type thing? If it is, then milk should be no problem (I have done this in the past)
This is an engineered enzyme. Should be a Nickel-loaded chelator attached to the HRP.
I know this question is rather old, but in case someone finds this useful, you can indeed block with 5% milk instead of BSA. I also do the incubation at 1:10000 in TBST without milk for more pure samples.
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