Description
Number of blots:
At least 20 blots, based on a 1 ?l/ml dilution and 5 ml diluted antibody per blot.
Important protocol notes:
1. The anti-mouse IgG for IP secondary antibody (HRP) detects mouse IgG antibodies (subtypes: IgG1, IgG2a, IgG2b, IgG3).
2. The anti-mouse IgG for IP secondary antibody (HRP) preferentially detects the non-reduced form of mouse IgG (IgG1, IgG2a, IgG2b, IgG3) over the reduced, SDS-denatured forms.
3. IP sample should be completely reduced/denatured before loaded onto a western blot.
4. Milk should be used as the blocking protein for the immunoblot.