Antibodies for Multiplex Immunofluorescence (mIF)

Multiplexed immunofluorescence (mIF) is a powerful technique for investigating complex biological systems and cellular interactions. It allows researchers to simultaneously detect multiple targets within a single tissue, providing a deeper understanding of cellular dynamics, spatial relationships, and biomarker expression patterns.

Table of Contents

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. 

Featured Multiplex IF Antibodies Qualified on COMET™

Figure 1: Detection of beta-Actin in Mouse Stomach viaseqIF™ staining on COMET™ using Rat Anti-Mouse beta-Actin Monoclonal Antibody (Catalog #MAB11702) Tissue was stained using the Alexa Fluor™ 555 Goat anti-Rat IgG Secondary Antibody. (Yellow; Lunaphore Catalog # DR555RT) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the cell membrane

Figure 2: Detection of CD21 in Human Lymph Node via seqIF™ staining on COMET™ using Rabbit Anti-Human CD21 Monoclonal Antibody (Catalog # NBP3-20354). Tissue was stained using the Alexa Fluor™ Plus 555 Goat anti-Rabbit IgG Secondary Antibody (Yellow; Lunaphore Catalog # DR555RB) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the membrane.

Figure 3: Detection of CD31 in Mouse Thymus via seqIF™ staining on COMET™ using Rat Anti-Mouse CD31 Monoclonal Antibody (Catalog # MAB11714). Tissue was stained using the Alexa Fluor™ 647 Goat anti-Rat IgG Secondary Antibody (Yellow; Lunaphore Catalog # DR647RT) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the membrane. 

COMET™ Qualified mIF Antibodies

AIF-1/Iba1alpha-Smooth Muscle Actin (SMA) - VerifiedAquaporin-4Arginase 1/ARG1
B7-1/CD80B7-H3Bcl-2beta-Actin
Beta-cateninBOB1CD11b (human) - VerifiedCD11b (mouse)
CD11c (human)CD11c (mouse)CD14CD163 -Verified
CD16a/Fc gamma Receptor IIIACD16b/Fc gamma Receptor IIIBCD19CD20
CD21CD3e (human)CD3e (mouse)CD31/PECAM-1 (human)
CD31/PECAM-1 (mouse) CD34 (human) - VerifiedCD34 (mouse)CD4 (human)
CD4 (mouse)CD44CD45 (human)CD45 (mouse)
CD45RACD68/SR-D1 (human)CD68/SR-D1 (mouse)CD69
CD73 -VerifiedCD8 (human)CD8 (mouse)c-Myc
Collagen I (human)Collagen I (mouse)Collagen IV -VerifiedCTLA-4
CXCL12/SDF-1CXCR7/RDC-1Cyclin D1Cytokeratin
DesminDPPIV/CD26E-Cadherin -VerifiedEpCAM/TROP-1 (human)
EpCAM/TROP-1 (mouse)ErbB2/Her2Fas Ligand/TNFSF6Fibrinogen
Fibroblast Activation Protein alpha/FAPFibronectinFOXP3GATA-3
Gelsolin/GSNGFAP (human)GFAP (mouse)Granzyme B
HLA-DRIDO-1IgDIgM
IRF4Kappa IgKi67LAG-3
Lambda IgMAP2 (human)MAP2 (mouse)MHC Class I/HLA-ABC
Na+/K+ ATPaseNCAM-1/CD56 (human)NCAM-1/CD56 (mouse)Neprilysin/CD10
P2Y12/P2RY12p53Pan-cadherin - VerifiedPax5/BSAP
PD-1PD-L1 (human)PD-L1 (mouse)Perforin
Phospho-ERK1/ERK2PodoplaninRBFOX3/NeuN (human)RBFOX3/NeuN (mouse)
S100A4 (mouse)S100BSLC22A2/OCT2SOX11
SOX2STING/TMEM173SOX2STING/TMEM173
VCAM-1/CD106VEGFR2/KDR/Flk-1 (mouse)VEGFR3/Flt-4 - VerifiedVimentin (human) - Verified
Vimentin (mouse)- VerifiedVISTA/B7-H5/PD-1HvWF-A2 

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

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

Multiplex IF (mIF) validated antibodies with a Lunaphore certified badge are taken through the qualification or verification process listed above. This rigorous testing process 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 Lunaphore Certified antibodies are also in the Lunaphore Panel Builder and added to the expanding COMET marker database.

Explore our curated selection of antibodies and take your spatial biology studies to new heights. All R&D Systems antibodies are provided in conjugation-ready format, free from BSA and Azide. Find our growing list of spatial biology tested antibodies from Novus Biologicals and R&D Systems below.

Have you used one of our antibodies in a spatial biology platform? Submit a review and receive a reward!

TargetPlatformCatalog NumberExternal Data LinkOMAP_ID
ABCA3Cell DIVE™NBP1-89310CitationOMAP-7
AGERCell DIVE™AF1145CitationOMAP-7
APP / β AmyloidNanostring GeoMx® DSPNBP2-13075AF647Data Image 
BST2 / CD317IMC (Hyperion™ and Hyperion Xti™)DDX0390P-100Citation 
CD11bNanostring GeoMx® DSPNB110-89474AF647Data Image 
CD161 / NK1.1IMC (Hyperion™ and Hyperion Xti™)NB100-77528Citation 
CD20Nanostring GeoMx® DSPNBP2-47840AF647Data Image 
CD20Nanostring GeoMx® DSPNBP2-47840AF594Data Image 
CD20Nanostring GeoMx® DSPNBP2-47840AF488Data Image 
CD3Nanostring GeoMx® DSPNBP2-54392AF647Data Image 
CD3Nanostring GeoMx® DSPNBP2-54392AF594Data Image 
CD3Nanostring GeoMx® DSPNBP2-54392AF532Data Image 
CD3Nanostring GeoMx® DSPNBP2-54392AF488Data Image 
CD4SIMSNBP2-25199Citation 
CD4Canopy CellScapeFAB8165GCitation 
CD68Nanostring GeoMx® DSPNBP2-34587AF488Data Image 
CD68Nanostring GeoMx® DSPNBP2-34587AF594Data Image 
CD68Nanostring GeoMx® DSPNBP2-34587AF647Data Image 
CD68Nanostring GeoMx® DSPNBP2-34587AF532Data Image 
CD69IMC (Hyperion™ and Hyperion Xti™)NBP1-51607Data Image 
CD7IMC (Hyperion™ and Hyperion Xti™)NBP2-37368Data Image 
CHGACODEXNBP2-34674Citation 1Citation 2OMAP-2, OMAP-6, OMAP-13
C-Kit / CD117Nanostring GeoMx® DSPNBP2-90037AF647Data Image 
CPS1SIMSNBP3-08970CitationOMAP-5
CTSLCell DIVE™NB100-1775CitationOMAP-7
CXCL13IBEXAF801Citation 1, Citation 2OMAP-1
CXCR6IBEXFAB2145RCitation 
ELA2 / Neutrophil ElastaseNanostring GeoMx® DSPMAB9167AF594Data Image 
ELA2 / Neutrophil ElastaseNanostring GeoMx® DSPMAB9167AF488Data Image 
FibronectinIBEXNBP2-22113AF532Citation 
GCGCODEXMAB12491CitationOMAP-6
GfapNanostring GeoMx® DSPNBP1-05197AF647Data Image 
GfapNanostring GeoMx® DSPNBP1-05197AF594Data Image 
GfapNanostring GeoMx® DSPNBP1-05197AF532Data Image 
GfapNanostring GeoMx® DSPNBP1-05197AF488Data Image 
GfapNanostring GeoMx® DSPNBP2-33184AF647Data Image 
GfapNanostring GeoMx® DSPNBP2-33184DL594Data Image 
GfapNanostring GeoMx® DSPNBP2-33184AF532Data Image 
GfapNanostring GeoMx® DSPNBP2-33184AF488Data Image 
GfapNanostring GeoMx® DSPNBP2-34413AF647Data Image 
GHRLCODEXMAB8200CitationOMAP-13
GP2CODEXNBP2-75781CitationOMAP-13
HAVCR1CODEXMAB1750CitationOMAP-9
HLA-GIMC (Hyperion™ and Hyperion Xti™)NB110-55297 OMAP-8
ITLN1CODEXAF4254CitationOMAP-2
KRT (Pan-Cytokeratin)Nanostring GeoMx® DSPNBP2-33200AF594Data Image 
KRT (Pan-Cytokeratin)Nanostring GeoMx® DSPNBP2-33200AF488Data Image 
KRT8CODEXNBP2-34501CitationOMAP-9
KRT8/18 (Cytokeratin)Nanostring GeoMx® DSPNBP2-34655AF594Data Image 
Laminin (Pan-Laminin)Nanostring GeoMx® DSPNB300-144AF532Data Image 
Laminin (Pan-Laminin)Nanostring GeoMx® DSPNB300-144AF488Data Image 
Laminin (Pan-Laminin)Nanostring GeoMx® DSPNB300-144AF594Data Image 
Laminin (Pan-Laminin)Nanostring GeoMx® DSPNB300-144AF647Data Image 
Lgr5Nanostring GeoMx® DSPNBP1-28904AF594Data Image 
LRP2CODEXMAB9578CitationOMAP-9
LUMIBEXAF2846Citation 1Citation 2OMAP-1, OMAP-11
LYVE1IBEX, Cell DIVE™, IMC (Hyperion™ and Hyperion Xti™), CODEXAF2089Citation 1Citation 2Citation 3Citation 4, Citation 5OMAP-1, OMAP-4OMAP-7OMAP-8OMAP-13
MMR/CD206IMC (Hyperion™ and Hyperion Xti™)AF2535Citation 
MRC1CODEXAF2534CitationOMAP-9
MUC1Nanostring GeoMx® DSPNBP2-47888AF594Data Image 
MUC2CODEXNB120-11197CitationOMAP-2
NefhNanostring GeoMx® DSPNB500-416AF594Data Image 
NEFHNanostring GeoMx® DSPNB500-416AF647Data Image 
NefhNanostring GeoMx® DSPNB500-416AF488Data Image 
NefhNanostring GeoMx® DSPNB500-416AF532Data Image 
NOS2Cell DIVE™NBP2-22119CitationOMAP-7
OCLNNanostring GeoMx® DSPNBP3-08879AF594Data Image 
OCLNNanostring GeoMx® DSPNBP3-08879AF647Data Image 
p53IBEXNB200-103PECitation 
PECAM1IMC (Hyperion™ and Hyperion Xti™)NB600-562 OMAP-8
Perilipin IMC (Hyperion™ and Hyperion Xti™)NB110-40760Data Image 
PODXLCODEXAF1658CitationOMAP-9
PPYCODEXMAB62971CitationOMAP-13
SPARCIBEXAF941Citation 1Citation 2OMAP-1, OMAP-11
SSTCODEXNBP2-37447Citation 1Citation 2OMAP-6OMAP-13
SYPCODEXNBP1-47483CitationOMAP-2
TGF-beta 1IMC (Hyperion™ and Hyperion Xti™)NBP1-80289Data Image 
UMODCODEXAF5144CitationOMAP-9

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