Description

Can Get Signal® immunostain is a reaction solution that contains an accelerator for antigen-antibody reactions, which improves sensitivity, specificity, and S/N of immunohistochemistry (IHC) and immunocytochemistry.

Features

  • Improves sensitivity, specificity, and S/N of IHC.
  • Can be applied to various detection systems (e.g., chromogenic, chemiluminescence, or fluorescence).
  • Can be used with ABC or polymer complex methods.
  • Solutions A and B exhibit various properties for improving results.
  • Can be used directly without dilution (Ready-to-use).

Applications

  • Immunohistochemistry (IHC)
  • Immunocytochemistry

Storage condition

Store at 4ºC

Components

This kit includes the following components. All reagents should be stored at 4ºC and protected from light.

Reagent Name Code No.
NKB-401 NKB-501 NKB-601
Solution A 5 ml 20 ml -
Solution B 5 ml - 20 ml

Typical reaction flow

Flow chart of immunostaining with Can Get Signal® immunostain

Application data

Example 1. Detection of PCNA using paraffin-embedded sections

The localization of PCNA (proliferating cell nuclear antigen) expression in human skin was detected using paraffin-embedded sections of the human skin tissue model TESTSKINTM (Toyobo). Detection was performed by the ABC method with anti-PCNA mouse monoclonal antibody as the primary antibody and biotinylated mouse IgG as the secondary antibody. Each antibody was diluted with Solution A of Can Get Signal® immunostain prior to use. As a control experiment, PBS(-) containing 1.5% normal horse serum (conventional method) was used instead of Solution A of Can Get Signal® immunostain. As a result, Can Get Signal® immunostain produced higher signals and lower background than the conventional method.

Fig. 1. IHC detection of PCNA using paraffin-embedded tissue sections

Example 2. Detection of paxillin with the fluorescent antibody method

The localization of paxillin in Swiss 3T3 cells was detected using anti-paxillin polyclonal antibody and Alexa488-conjugated rabbit IgG antibody. As a result, the exposure time for detection could be reduced from 3 s to 1 s by using Can Get Signal immunostain®, and the detailed fiber structure (fibrillar adhesion) could also be detected by using Can Get Signal® immunostain. However, the structure could not be detected with the conventional method due to high background signals.
Fig. 2C shows a merged picture of the immunological detection with anti-paxillin and anti-phospho-tyrosine antibodies, and actin staining.

Fig. 2. Immunocytochemistry detection of paxillin with the fluorescent antibody method

*The data was kindly provided by Dr. Harada, Tokyo Institute of Technology.