My cloned gene contain His-C terminal tag. I would like to check my protein using western blot. So could you please advice me what sort of antibody could be use. I prefered to do the detection with Alkaline phosphatase using NBT and BCIP as the substrate. Do I need to design a primary antibody for my protein or can I use Anti-His(C-term)-AP Antibody directly?
You might use a polyclonal antibody against your protein in conjunction with the commercial anti His tag antibody in case the His tag is cleaved
Are you referring the primary ab or secondary ab or a common ab for his tag which no need primary or secondary abs ?Please explain I am confuse, do I need a primary ab to be synthesised my self using my protein if I already has a his - tag at treminal?
Primary antibodies against the his tag are commercially available. You can use this and use a secondary antibody thats conjugated to HRP for chemiluminescent detection of ur his tagged protein.
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