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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
Beta-CateninHuman, Mouse, RatMAB1329 
CadherinHuman, MouseMAB18385
CD11b/Integrin alpha MHumanMAB16992
CD163HumanNB110-40686
CD20HumanMAB42252
CD34HumanNBP2-32932
CD4HumanMAB11560
CD45HumanMAB14303
CD45RAHumanMAB11444
CD68/SR-D1HumanMAB2040 
CD73HumanNBP2-25237
CD8 alphaHumanMAB3801
Collagen IVHuman, MouseNBP3-25416
CXCR7/RDC-1HumanMAB4227 
EpCAM/TROP-1HumanMAB960 
FoxP3Human, MouseMAB8214 
GATA-3Human, MouseMAB6330 
Ki67/MKI67HumanMAB7617 
LAG-3HumanNBP2-80059
S100BHumanMAB18203
Smooth Muscle ActinMulti-speciesNBP2-33006
VimentinHuman, Mouse, RatMAB2105 

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.