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. Author manuscript; available in PMC: 2022 Jan 1.
Published in final edited form as: Vet Pathol. 2020 Nov 19;58(1):181–204. doi: 10.1177/0300985820970144

Table 4.

Immunohistochemistry methods for antibodies used in the study

Antigen Speciesa Dilution Source, Catalog No. Platform Antigen Retrieval Visualization
hNuMA1 H 1:75 Lifespan Biosciences, LS-B11047 Ventanab HIER, CC2c OmniMapd
CD117 /c-Kit H/M 1:200 Dako Agilent, A4502 Ventana HIER, CC1e IVIEWf
MCT H/M 1:6000 Abcam, ab151757 Ventana HIER, CC2 OmniMap
hMBP H 1:20 Bio-Rad, MCA5751 Ventana HIER, CC2 OmniMap
mMBP M 1:3000 Mayo, MT-14.7 Biocareg EIERh Bio polyi
hCD68 H 1:50 Dako Agilent, M0876 Ventana HIER, CC1 IVIEW
hCD163 H 1:50 Cell Marque, MRQ-26 Ventana HIER, CC1 IVIEW
F4/80 M 1:500 Invitrogen, MF48000 Biocare HIER, TRj Biotinylk
hCD45 H RTUl Ventana, 760-2505 Ventana HIER, CC1 OmniMap
CD3 H/M 1:1000 Santa Cruz, sc-1127 Ventana HIER, CC1 OmniMap
PAX5 H/M 1:1000 Abcam, ab109443 Leicam HIER, ER2n BPRDo
hCD4 H RTU Leica, PA0427 Leica HIER, ER2 BPRD
hCD8 H RTU Leica, PA0183 Leica HIER, ER1p BPRD
GFP N/A 1:2000 CLONTECH, 632381 Ventana HIER, CC2 Purpleq
GATA1 H/M 1:4000 Abcam, ab131456 Ventana HIER, CC1 OmniMap
MPO H/M 1:1200 Dako, A0398 Ventana HIER, CC1 OmniMap

Abbreviations: MCT, mast cell tryptase; MBP, major basic protein; GFP, green fluorescent protein; MPO, myeloperoxidase

a

Species reactivity; H, human; M, murine; N/A, not applicable

b

Ventana DISCOVERY ULTRA automated stainer, Ventana Medical Systems, Inc, Tucson, AZ

c

Heat-induced epitope retrieval, Cell conditioning media 2, Ventana Medical Systems, Inc

d

DISCOVERY OmniMap anti-rabbit HRP (760-4311), DISCOVERY ChromoMap DAB kit (760-159); Counterstain Hematoxylin II (Roche, Indianapolis, IN, 790-2208) and Blueing reagent (Roche, 760-2037)

e

Heat-induced epitope retrieval, Cell conditioning media 1, Ventana Medical Systems, Inc

f

IVIEW DAB Kit (760-091); Counterstain Hematoxylin II (Roche, Indianapolis, IN, 790-2208) and Blueing reagent (Roche, 760-2037)

g

intelliPATH FLX, BioCare Medical, LLC, Pacheco, CA

h

Enzyme-induced epitope retrieval with Proteinase K, DAKO S3020

i

Rabbit anti-rat secondary antibody (Vector Labs, BA-4001), Rabbit-on-Rodent AP-Polymer (BioCare Medical, Concord, CA, RMR625H), and Fast Red chromogen kit (BioCare Medical, IPK5017G80); Counterstain hematoxylin (BioCare Medical, IPCS5006L)

j

HIER with Decloaking Chamber NsGEN and Target Retrieval, pH 6

k

Biotinylated rabbit anti-rat IgG, mouse absorbed secondary antibody (Vector Labs, BA-4001); Lab Vision streptavidin peroxidase (Thermo Scientific, TS-125-HR); DAB chromogen kit (Thermo Scientific, TA-125-QHDX); Counterstain hematoxylin (BioCare Medical, IPCS5006L)

l

RTU, ready to use

m

Leica BOND-MAX automated stainer, Leica Biosystems, Buffalo Grove, IL

n

HIER with Bond Epitope Retrieval Solution 2 (ER2)

o

Bond Polymer Refine Detection (DS9800); Counterstain Hematoxylin Refine Kit (Leica Biosystems, DS9800)

p

HIER with Bond Epitope Retrieval Solution 1 (ER1)

q

DISCOVERY OmniMap anti-rabbit HRP (760-4311), DISCOVERY ChromoMap Purple kit (760-229); Counterstain Hematoxylin II (Roche, Indianapolis, IN, 790-2208) and Blueing reagent (Roche, 760-2037)