Multiplex Immunofluorescence (mIF) on COMET™

The COMET™ platform, from Lunaphore, a Bio-Techne brand, is a fully-automated, high-throughput, spatial biology platform that performs sequential immunofluorescence (seqIF™) through staining, imaging, and elution cycles. It offers the unique flexibility to work with off-the-shelf, non-conjugated primary antibodies. 

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Antibody Testing Process on COMET™

Bio-Techne's antibody validation experts have collaborated and trained alongside Lunaphore's research and development team covering protocols and requirements involved in the antibody validation process on COMET™. This includes the antibody qualification process for protocol optimization, with a focus on assessing specificity, sensitivity, and elution efficiency using positive control tissue. At Bio-Techne, our IHC validated antibodies have already undergone a rigorous process to be added to the catalog.  Antibody qualification process for COMET™ involves several key aspects as shown in Figure 1.

Illustration showing antibody testing process for mIF on COMET with descriptions of antibody sensitivity and specificity

Figure 1. Steps of Antibody Testing Process on COMET™ that leads to Lunaphore Certified badge

You’ll find a Lunaphore Certified badge on our mIF validated antibody images. This ensures that antibodies, from R&D Systems™ and Novus Biologicals™, part of Bio-Techne, deliver high-quality and specific staining on the COMET™ platform.  Bio-Techne mIF validated antibodies are also in the Lunaphore Panel Builder and added to the expanding COMET™ marker database.

COMET™ Qualified mIF Antibodies

TargetSpecies ReactivityCatalog Number
B7H3HumanMAB11611
CD10/NeprilysinHumanMAB11821
CD3HumanMAB10670
CD34HumanMAB72273
CD44HumanMAB7045
CytokeratinMulti-speciesNBP2-29429
DesminHumanMAB11607
E-cadherinHuman, Mouse, MonkeyNBP2-19051
ErbB2/Her2HumanMAB11291
Fas Ligand/TNFSF6HumanMAB0951
FibronectinHumanMAB19182
GelsolinHumanMAB8170
GFAPHumanMAB25941
IgDHumanMAB9857
Kappa IgHumanMAB100501
Lambda IgHumanMAB10049
MAP2HumanMAB8304
PAX5HumanMAB3487
SLC22A2/OCT2HumanMAB6547
SOX2HumanMAB2018

Table 1: List of COMET™ Qualified mIF Antibodies from R&D Systems™ and Novus Biologicals™ part of Bio-Techne.

Featured Multiplex IF Antibodies qualified on COMET™

Detection of Cadherin in human colon cancer via Multiplex Immunofluorescence on COMET™

Figure 2: Detection of Cadherin in Human Colon Cancer via Multiplex Immunofluorescence staining on COMET™ using Mouse Anti-Human/Mouse Cadherin Pan Specific Monoclonal Antibody (R&D Systems, Catalog# MAB18385). Tissue was stained using the Alexa Fluor™ 647 Goat anti-Mouse IgG Secondary Antibody (Yellow; Lunaphore Catalog # DR647MS) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the membrane.

Detection of CD4 in Human Tonsil via Multiplex Immunofluorescence (mIF) staining on COMET™"

Figure 3: Detection of CD34 in Human Kidney via Multiplex Immunofluorescence staining on COMET™ using Mouse Anti-Human CD34 Monoclonal Antibody (Novus Biologicals, Catalog # NBP2-32932). Tissue was stained using the Alexa Fluor™ Plus 647 Goat anti-Mouse IgG Secondary Antibody (Yellow; Lunaphore Catalog # DR647MS) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the cell membrane.

Detection of CD34 in Human Kidney via Multiplex Immunofluorescence (mIF) staining on COMET™

Figure 4: Detection of CD4 in Human Tonsil via Multiplex Immunofluorescence staining on COMET™ using Rabbit Anti-Human CD4 Monoclonal Antibody (R&D Systems, Catalog # MAB11560). Tissue was stained using the Alexa Fluor™ Plus 647 Goat anti-Rabbit IgG Secondary Antibody (Yellow; Lunaphore Catalog # DR647RB) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the membrane.

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