How does the Clean-Blot IP Detection Kit work?
This assay uses a HRP conjugate or AP conjugate that binds to native IgG from various host species, allowing clear, specific Western blot detection from IP experiments without interference from denatured IgG. It is used simply as a substitute for secondary antibodies.
Does the Clean-Blot IP Detection Reagent bind to all antibody classes and subclasses?
The reagent binds to IgG from a wide range of species:
bovine IgG2, goat IgG2, human IgG1, IgG2, IgG4, mouse IgG2a, IgG2b, IgG3, rat IgG2c, sheep IgG2, pig, dog and cat. It will not bind to rat IgG2a and IgG2b or mouse IgG1. If in doubt whether this detection reagent will bind to a specific antibody, perform a dot-blot analysis before the experiment.
What blocking buffers are compatible with this detection reagent?
The Clean-Blot IP Detection is compatible with bovine serum albumin, SuperBlock and StartingBlock Blocking Buffers and 5% nonfat milk. Verify compatibility with other blocking buffers by dot-blot analysis.
What substrates are compatible with this detection reagent?
For best results, use the the Clean-Blot IP Detection with SuperSignal Substrates or Pierce ECL Western Blotting Substrate. When using other HRP or AP substrates, empirically determine the optimal concentration of the Clean-Blot IP Detection Reagent.
Can I strip and reprobe my blot after using this detection reagent?
Yes. When using a chemiluminescent substrate, membranes can be stripped and reprobed similar to probing with secondary antibodies.