2-step Plus Poly-HRP Anti Rabbit/Mouse IgG Detection System

2-step plus is a two-step immunohistochemical broad spectrum detection reagent, recently launched by the Elabscience company. It polymerizes monovalent Fab fragments of secondary antibody and enzyme, which replaces the traditional method through secondary antibody and tertiary antibody binding signal. This method not only retains specific binding ability of antibody with antigen, but can also effectively avoid steric hindrance caused by excessive polymer molecules. Compared with the traditional SP three-step method, this kit has the characteristics of being simple, rapid and sensitive. The detection system no longer uses biotin, so it eliminates background staining by endogenous biotin. It can be used to detect monoclonal or polyclonal antibodies of rabbit/mouse source. The unique polymer auxiliary agent for macromolecular detection system achieves better primary antibody binding for detection. This detection system is now the most sensitive immunohistochemical detection system available from Elabscience.

Components

IHC Detection Procedure

1. Section paraffin,dewax conventionally
2. According to special requirements of applied primary antibody, pretreat the tissue section;
3. Incubate with 3% H2O2 deionized water for 10 min to eliminate endogenous peroxidase activity;
4. Wash with PBS or TBS,v;
5. Add primary antibody with proper dilution ratio, incubate at 20~37℃ for 1~2h or at 4℃ for one night (then rewarming at 37℃ for 30min), wash with PBS or TBS, v;
6. Add reagent 1, incubate at room temperature or 37℃ for 20min, wash with PBS or TBS, 2 min, 3 times;
7. Add reagent 2, incubate at room temperature or 37℃ for 20~30min, wash with PBS or TBS, c;
8. Color development with DAB;
9. Wash with deionized water;
10. Make the section counterstained, dewatered, transparent if the experiment requires;
11. Mount the section with proper mounting medium and then observe.

Storage

Stored at 2°C – 8°C. Shading light.

Manual and Datasheet