Do I use Direct or Indirect labelling (antibodies)?
Gold Conjugates may be used directly or indirectly to label antigens as already indicated. In the direct method the gold is conjugated to primary antibody, whether monoclonal or polyclonal. This allows a single incubation to be performed and provides the simplest detection system. In the indirect method a primary unlabelled antibody is applied to the spectrum to locate the antigen. This is followed by the gold secondary antibody that detects the primary antibody. This gold labelled secondary antibody is almost always affinity purified polyclonal. An extension of the indirect method may be used where there is no appropriate gold labelled secondary to match the primary antibody. For example, if the unlabelled primary is from Pig and there is no matching gold labelled anti-Pig then an unlabelled anti-Pig (sat Rabbit anti-Pig) may be used as a secondary step followed by a gold labelled third antibody (say Goat anti-Rabbit) that will detect the second antibody. The indirect method is the mo
Related Questions
- I have been granted confidentiality for one of my substances. Can this decision be revisited in the future if the classification and labelling of this substance changes?
- Can I fix my cells after staining with eFluorTM Nanocrystal conjugated antibodies?
- Is there evidence of labelling that ABI has done on avian antibodies?
