Multiplex IHC kit

Multiplex tyramide signal amplification on frozen tissue sections
The StreptaClick–HRP Multiplex IHC kit empowers you to perform multiplex immunostaining with tyramide signal amplification (TSA) on frozen tissue sections. Other TSA kits on the market can only be used for formalin fixed paraffin embedded sections (FFPE).

Exceptional signals on frozen tissue sections
The kit results in 10-50 times signal amplification compared with directly conjugated antibodies.

Click chemistry conjugation of HRP
The StreptaClick–HRP reagent in the kit attaches with a click reaction horseradish peroxidase (HRP)  to each biotinylated antibody that will be used in the staining. The HRP–labeled biotinylated antibodies are then added in cycles to the tissue and developed with TSA (TSA reagents are not provided in the kit).

No boiling in microwave oven
The kit contains an HRP block buffer optimized for preserving the morphology of frozen tissue sections. The HRP block buffer rapidly quenches the HRP enzymatic activity after each antibody cycle. Hence, there is no need for antibody removal with heat treatment.

7-color TSA multiplex HC in one day
The staining protocol is much faster compared to standard TSA protocols, since it does not include secondary antibodies or boiling in microwave oven. The fast protocol allows 7-color TSA multiplex IHC (DAPI + 6 biotinylated antibodies) to be completed in one day! For direct imaging of the multistained tissue we recommend using a microscope equipped with SpectraSplit.

Bright signals. Comparison in exposure time and image quality in pictures captured from frozen tissue sections stained with biotinylated antibody + StreptaClick-HRP Multiplex IHC kit (left panel, exposure time 50 ms, x10 objective) or directly conjugated antibody (right panel, exposure time 700 ms, x10 objective). Antibodies: biotin-conjugated anti-CD4, clone GK1.5, dilution 1:400 (1.25 µg/ml) and AlexaFluor488-conjugated anti-CD4, clone GK1.5, dilution 1:100 (5 µg/ml). Tissue: mouse spleen.

Preserved morphology. Preserved morphology of frozen tissue sections after five TSA cycles using the StreptaClick-HRP Multiplex IHC kit. Macrophages in mouse spleen were immunostained with biotinylated antibody against F4/80 (clone BM8) and StreptaClick-HRP Multiplex IHC kit. Left panel: Section stained with anti-F4/80 at cycle 1. Right panel: Section stained with anti-F4/80 at cycle 5. Fluorochrome: tyramide CF594.

Efficient block of enzymatic activity by HRP block buffer. Two frozen tissue sections from mouse spleen were incubated with biotinylated antibody against F4/80 (clone BM8) labeled with StreptaClick-HRP. Before color development, one section was treated 12 minutes with HRP block buffer (left panel), while the control section was not treated (right panel). Both sections were then incubated 10 minutes with tyramide CF555 to develop color.

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