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SignalStar Multiplex IHC Panel Builder

Before You Get Started…

SignalStar Multiplex Immunohistochemistry (mIHC) is a cutting-edge technology that uses specific primary antibodies and fluorescent detection reagents to flexibly and reliably label cellular proteins in FFPE tissue samples. The SignalStar technology from CST enables you to quantify multiple biomarkers simultaneously with fully customizable, highly-validated antibody panels—accelerating data generation and letting you focus on results.

The SignalStar Multiplex IHC Panel Builder provides a seamless, effortless multiplex IHC panel design. In four basic steps, you can quickly and easily design your experiment by choosing your target species, antibodies, fluorophores, and imaging sequence. Or you can choose your species and antibodies, and let the automated panel builder guide you the rest of the way.

How SignalStar Multiplex IHC Works

SignalStar Multiplex IHC WorkFlow Diagram

Figure Legend: The SignalStar assay allows for the simultaneous detection of up to 8 targets in formalin-fixed paraffin embedded (FFPE) tissue. Deparaffinized and rehydrated FFPE tissue sections undergo antigen retrieval (A), and all antibodies in your plex size of choice (3-8 maximum unique oligo-conjugated antibodies) are added in one primary incubation step (B). Complementary oligonucleotides with fluorescent dyes (channels: 488, 594, 647, 750 nm) amplify the signal of up to 4 oligo-conjugated antibodies in the first round of imaging by building large oligo branched structures off the antibody (C-D). If the plex size is greater than 4, the first round of oligonucleotides and fluorophores are gently removed (E), and a second round is added to amplify the signal of up to 4 additional oligo-conjugated antibodies (F). The two images are then aligned computationally with either proprietary or open-source software to generate the full, up to 8-plex image (G).

How to Use the SignalStar Multiplex IHC Panel Builder

Step 1: Choose Your Target Species

SignalStar Multiplex IHC antibodies are validated to confirm specific reactivity with the species indicated. Not all antibodies work in all species.

Step 2: Choose Your Antibodies

Choose 3-8 antibodies validated for your target of interest. Like all CST® antibodies, the ones identified for use in your SignalStar panels have each been individually validated for use in their intended application. So like our other antibodies, they're guaranteed to perform as expected—and to provide you with reliable, reproducible results, the first time and every time.

Step 3: View/Edit Your Panel Builder Results

The SignalStar Multiplex IHC Panel Builder has the unique capability to automate design of your multiplex IHC panel, pairing antibodies to fluorophores and imaging rounds with inherent accuracy and efficiency. You can review and confirm the panel builder results, or choose to edit for more control of your experimental design.

Considerations when designing your own Multiplex IHC Panel

Step 4: Confirm Your Order

All antibodies and buffer kits needed for your experiment are automatically added to your cart. You can continue to add panels in the SignalStar Multiplex IHC Panel Builder multiple times. Just be careful not to remove components of prior panels once they're in your basket.


Need Support?

We provide expert support to optimize the application of SignalStar Multiplex IHC technology to reduce the time in your research. Our team is available to answer questions and offer technical assistance.

Contact our SignalStar group directly at [email protected].

For additional support, visit the Learn & Support page.