Split peroxidases and methods of use
An imaging method utilizing a split peroxidase is described herein. Imaging methods involve contacting a cell with a split peroxidase and a substrate thereof to allow conversion of a substrate into a product via an enzymatic reaction catalyzed by the reconstitute split peroxidase. Also disclosed herein are split peroxidases, related products and kits.
1. An imaging method, comprising: providing a sample containing a cell that expresses a split peroxidase comprising two or more separate components of a peroxidase, and contacting the sample with a peroxidase substrate to allow conversion of the peroxidase substrate into a product via an enzymatic reaction catalyzed by a reconstituted peroxidase that forms when the two or more components of the split peroxidase interact, wherein the product releases a detectable signal,
wherein said split peroxidases are capable of performing any of the following: converting said product without any assistance from additional protein-protein interaction, enduring chemical fixation, ultra-sensitive synapse detection, achieving greater sensitivity than split-GFP constructs or converting said product with assistance from a protein-protein interaction.
2. The method of claim 1 , wherein the signal is detectable by a microscope.
3. The method of claim 2 , wherein the signal is detected by electron microscopy.
4. The method of claim 2 , wherein the signal is detected by fluorescence microscopy.
5. The method of claim 1 , wherein the signal is detectable by a chemiluminescence.
6. The method of claim 1 , wherein the split peroxidase is a split horse radish peroxidase (HRP).
7. The method of claim 1 , wherein the split peroxidase is a split ascorbate peroxidase (APX).