This kit contains 3 TSA dyes for high sensitivity multiplex IHC.
Applications
IHC, ICC/IF
Principle of Assay
Tyramide signal amplification (TSA) is a highly sensitive fluorescent method for amplifying immunohistochemical staining when using HRP-conjugated antibodies. After applying HRP-labeled antibody to the sample and washing as in a normal IHC workflow, the tyramide fluorochrome is diluted in tyramide amplification buffer and applied to the samples for 10 minutes to develop the fluorescence. This is then repeated for multiple HRP-labeled antibodies by quenching HRP (HRP-block provided in StreptaClick® HRP Multiplex IHC Kit) and repeating the process for each antibody in sequence.
Shipped at +4°C. Store in the dark at -20°C. Before use, dissolve each vial in 100µl DMSO + 100µl PBS. Aliquot in vials and store at -20°C. The aliquots can be stored in the freezer for at least 12 months. Once thawed, store at +4°C and use within a week.
General Notes
The TSA 3 color kit includes the following fluorophores: Tyramide 488, Tyramide 555, and Tyramide 647. This kit is designed to be used in combination with our StreptaClick® HRP Multiplex IHC Kit. This kit contains sufficient reagents for 250 stainings, assuming 100µl per staining.
Disclaimer
This product is for research use only. It is not intended for diagnostic or therapeutic use.
Immunohistochemistry - Tyramide Signal Amplification Kit - 3 Color (A329042)
Four-color TSA immunohistochemistry of PFA-fixed, frozen mouse small intestine section using antibodies that have been biotinylated and labelled with the StreptaClick® HRP Complete Tyramide Signal Amplification and Biotinylation Kit - 5 Color. Tissue was mounted in OCT compound and frozen in liquid nitrogen before cutting 10 µm sections. The frozen tissue section was fixed in acetone (30 seconds in ice cold 50% acetone/50%water + 10 min in ice cold 100% acetone), air dried, and washed in PBS. Immunohistochemistry was performed using the following biotinylated antibodies: biotin rat anti-CD31 [MEC13.3] (Tyramide 488; green), biotin mouse anti-human alpha actin [1A4] (Tyramide 594; red), biotin rat anti-CD326 [G8.8] (Tyramide 555; yellow), biotin rat anti-mouse CD8a [53-6.7] (Tyramide 647; blue).
Immunohistochemistry - Tyramide Signal Amplification Kit - 3 Color (A329042)
Five-color TSA immunohistochemistry of PFA-fixed, frozen mouse spleen section using antibodies that have been biotinylated and labelled with the StreptaClick® HRP Complete Tyramide Signal Amplification and Biotinylation Kit - 5 Color. Tissue was fixed in 4% paraformaldehyde/10% sucrose for 30 min at +20°C, followed by incubation in 30% sucrose at +4°C overnight before freezing and cutting 10 µm sections. The frozen tissue section was fixed in acetone at room temperature for 10 min, air dried, and washed in PBS. Immunohistochemistry was performed using biotinylated antibodies and TSA dyes for color development: biotin rat anti-mouse CD4 [GK1.5] (Tyramide 430; cyan), biotin hamster anti-mouse CD11c [N418] (Tyramide 488; green), biotin rat anti-mouse B220 [RA3-6B2] (Tyramide 555; yellow), biotin rat anti-mouse CD8a [53-6.7] (Tyramide 594; red), and biotin rat anti-mouse F4/80 [M1/70] (Tyramide 647; magenta).
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