Multiplex Immunohistochemistry

Location: Thomas Building, DE-360
Contact phone: (206) 667-6166
Contact fax: (206) 667-6845
Contact e-mail: exphisto@fredhutch.org

Multiplex IHC allows investigators to characterize the tumor microenvironment (TME) by staining up to six biomarkers on one tissue section.  Experimental Histopathology (EH) staff use automated OPAL (PerkinElmer) workflow on the Leica Bond Rx staining platform.  Using this method the best antibodies can be selected based on performance with no need to worry about cross-reactivity.  Single cell co-expression can be confirmed while retaining morphological context and conserving precious tissue samples. 

Multiplex image acquisition using the Perkin Elmer Vectra 3.0 Automated Imaging Platform is a collaborative effort coordinated between EH staff and investigators. Images obtained in this way can be used in conjunction with HALO Multiplex and Highplex Image Analysis Modules to analyze on a cell-by cell basis, selecting regions of interest throughout tissue sections.

The following is a list of antibodies available. Use the specific mIHC form to request service.

HUMAN PRIMARY ANTIBODIES
Antibody Clone Manufacturer/#
AE1/AE3 AE1/AE3  DAKO / M3515
Arginase-1 EP261 BioSB / BSB2451
B7H3/CD276 RBT B7H3 BioSB / BSB2814
Bcl-6 EP278  Bio SB / BSB2820
CCR5 MC-5 Mack
CD11b Ep45 BioSB / BSB6440
CD11c EP157 BioSB / BSB6447
CD14  EP128 BioSB / BSB6460
CD14  EPR3653 Cell Marque/ 114R-14
CD16 (FcyRIII) SP175 Abcam / ab183354
CD16 (FcyRIII) EP364 BioSB / BSB3324
CD163 EP324 BioSB/ BSB3276
CD19 60MPO31 Affymetrix / 14-0194
CD20 L26 BioSB / BSB5194
CD20 L26 DAKO / M0755
CD206 Polyclonal Novus / NBP1-90020
CD3 RBT-CD3e BioSB / BSB5145
CD3 SP7 Thermo / RM-9107
CD3 CD3-12 BioRad / MCA1477
CD31 JC70A DAKO / M0823
CD33 RBT-CD33 BioSB / BSB3451
CD33 (Siglec-3) 44M12D3 Novus / NBP2-22377
CD33 (Siglec-3) 2C6B7 Novus / NBP2-37388
CD34 QBEnd10 Dako / M7165
CD4 SP35 Cell Marque/ 104R-16
CD4 RBt-CD4 BioSB / BSB5152
CD40L D5J9Y Cell Signal / 15094S
CD45 LCA 2B11+PD7/26 DAKO / M0701
CD45RA 4KB5 BioSB / BSB5257
CD45RO UCHL-1 BioSB / BSB5264
CD56 123C3.D5 BioSB /BSB5271
CD57 BSB-10 BioSB /BSB5278
CD66b G10F5 BD Bio/ 555723
CD68 PG-M1 DAKO/ M0876
CD68 BSB-8 BioSB / BSB5292
CD8 EP334 BioSB / BSB2849
CD8 144B DAKO / M7103 
CK20 Ks20.8 DAKO / M7019
CK20 Ks20.8 BioSB / BSB5390
CMET 4AT44 Millipore / MAB3729
COX-2 RBT-COX2 BioSB / BSB5362
CTLA-4 BSB-88 BioSB / BSB2884
FoxP3 236A/E7 eBiosc / 14-4777-82
FoxP3 EP340 BioSB / BSB2926
GATA-3 L50-823 BioSB/ BSB2674
GzB EP230 BioSB / BSB2409
HLA-DR  EP96 BioSB / BSB6797
HLA-DR  TAL.1B5 DAKO / M0746
HLA-DR B1 β-Chain EP191 BioSB / BSB2437
ICOS SP98 Novus / NBP2-12499
IgG4 EP138 BioSB / BSB6817
IL-18RA Poly R&D / AF870
Ki67 MIB1 DAKO / M7240
MHC-I (HLA-A,B,C) EMR8-5 MBL / D226-3
PD-1 D4W2J Cell Signaling / 86163
PD-1 NAT-105 BioSB/ BSB6216
PD-L1  E1L3N Cell Signaling / 13684
PD-L1/ CD274 RBT-PDL1 BioSB/ BSB2653
S100A9 Ep185 BioSB / BSB2172
Sox10 Polyclonal CellMarque / 383A-74
T-Bet/TBX-2 EP263 BioSB / BSB2605
TIM-3 D5D5R Cell Signaling / 45208S
     
MOUSE PRIMARY ANTIBODIES
Antibody Clone Manufacturer /#
CD3 CD3-12 BioRad / MCA1477
CD4 4SM95 Thermo / 14-9766-32
CD45 LCA 30-F11 BD Pharm / 550539
CD45R B220 RA3-6B2 Chemicon / CBL1342
CD8a 4SM15 Thermo / 14-0808-82
F4/80 CI:A3-1 BioRad / MCA497GA
GFP Polyclonal Thermo / A11122
Sox10 Polyclonal CellMarque / 383A-74