Western blot detection of IP proteins typically has high background resulting from the use of the same primary antibody in both the IP and the Western blot. This background is reduced using the Pure-IPTM Western Blot Detection Kit by using a proprietary HRP Conjugate for detection of the primary antibody. The conjugate is specific to properly folded antibodies and therefore is unable to detect the contaminating denatured heavy and light chains present on the membrane.
The Pure-IPTM Western Blot Detection Kit is compatible with existing protocols for sample preparation, IP, SDS-PAGE, and transfer. After transfer, membranes are blocked with the kit Pure-IPTM Blocking Solution and then probed with a primary antibody diluted with Pure-IPTM Blocking Solution. An HRP conjugate is then used for detection in place of a secondary antibody.