Table 1.

Triple fluorescence staining panel used for immunohistochemistry

Targets
 Vimentin IST1 CD68 Nucleus 
Isotype (primary ab) Rabbit polyclonal (ab92547; Abcam)
(1:250 dilution) 
Mouse IgG2a (66989-1-Ig; Proteintech)
(1:750 dilution) 
Mouse IgG3 (M0876; DAKO)
(1:100 dilution) 
 
Secondary ab Donkey anti-rabbit IgG (ab150075; Abcam)
(1:50 dilution) 
Rat anti-mouse IgG2a (RMG2a-62; BioLegend)
(1:20 dilution) 
Goat anti-mouse IgG3 (1101-01; Southern bio)
(1:75 dilution) 
 
Tertiary ab  Donkey anti-rat IgG (ab150153; Abcam)
(1:40 dilution) 
Donkey anti-goat IgG (ab175704; Abcam)
(1:75 dilution) 
 
Conjugate/dye AF647 AF488 AF568 DAPI 
Longpass filter 610LP 515LP 590LP 420LP 
 Targets 
 CD18 IST1 CD68 Nucleus 
Isotype (primary ab) Rabbit polyclonal (a2173; Abclonal)
(1:150 dilution) 
Mouse IgG2a (66989-1-Ig; Proteintech)
(1:750 dilution) 
Mouse IgG3 (M0876; DAKO)
(1:100 dilution) 
 
Secondary ab Donkey anti-rabbit IgG (ab150075; Abcam)
(1:50 dilution) 
Rat anti-mouse IgG2a (RMG2a-62; BioLegend)
(1:20 dilution) 
Goat anti-mouse IgG3 (1101-01; Southern bio)
(1:100 dilution) 
 
Tertiary ab  Donkey anti-rat IgG (ab150153; Abcam)
(1:40 dilution) 
Donkey anti-goat IgG (ab175704; Abcam)
(1:100 dilution) 
 
Conjugate/dye AF647 AF488 AF568 DAPI 
Longpass filter 610LP 515LP 590LP 420LP 
 Targets 
 CD18 IST1 Vimentin Nucleus 
Isotype (primary ab) Rabbit polyclonal (a2173; Abclonal)
(1:150 dilution) 
Mouse IgG2a (66989-1-Ig; Proteintech)
(1:750 dilution) 
Mouse IgG1 (RV202; Santa Cruz)
(1:300 dilution) 
 
Secondary ab Donkey anti-rabbit IgG (ab150075; Abcam)
(1:50 dilution) 
Rat anti-mouse IgG2a (RMG2a-62; BioLegend)
(1:20 dilution) 
Goat anti-mouse IgG1 (1071-01; Southern bio)
(1:100 dilution) 
 
Tertiary ab  Donkey anti-rat IgG (ab150153; Abcam)
(1:40 dilution) 
Donkey anti-goat IgG (ab175704; Abcam)
(1:100 dilution) 
 
Conjugate/dye AF647 AF488 AF568 DAPI 
Longpass filter 610LP 515LP 590LP 420LP 
Targets
 Vimentin IST1 CD68 Nucleus 
Isotype (primary ab) Rabbit polyclonal (ab92547; Abcam)
(1:250 dilution) 
Mouse IgG2a (66989-1-Ig; Proteintech)
(1:750 dilution) 
Mouse IgG3 (M0876; DAKO)
(1:100 dilution) 
 
Secondary ab Donkey anti-rabbit IgG (ab150075; Abcam)
(1:50 dilution) 
Rat anti-mouse IgG2a (RMG2a-62; BioLegend)
(1:20 dilution) 
Goat anti-mouse IgG3 (1101-01; Southern bio)
(1:75 dilution) 
 
Tertiary ab  Donkey anti-rat IgG (ab150153; Abcam)
(1:40 dilution) 
Donkey anti-goat IgG (ab175704; Abcam)
(1:75 dilution) 
 
Conjugate/dye AF647 AF488 AF568 DAPI 
Longpass filter 610LP 515LP 590LP 420LP 
 Targets 
 CD18 IST1 CD68 Nucleus 
Isotype (primary ab) Rabbit polyclonal (a2173; Abclonal)
(1:150 dilution) 
Mouse IgG2a (66989-1-Ig; Proteintech)
(1:750 dilution) 
Mouse IgG3 (M0876; DAKO)
(1:100 dilution) 
 
Secondary ab Donkey anti-rabbit IgG (ab150075; Abcam)
(1:50 dilution) 
Rat anti-mouse IgG2a (RMG2a-62; BioLegend)
(1:20 dilution) 
Goat anti-mouse IgG3 (1101-01; Southern bio)
(1:100 dilution) 
 
Tertiary ab  Donkey anti-rat IgG (ab150153; Abcam)
(1:40 dilution) 
Donkey anti-goat IgG (ab175704; Abcam)
(1:100 dilution) 
 
Conjugate/dye AF647 AF488 AF568 DAPI 
Longpass filter 610LP 515LP 590LP 420LP 
 Targets 
 CD18 IST1 Vimentin Nucleus 
Isotype (primary ab) Rabbit polyclonal (a2173; Abclonal)
(1:150 dilution) 
Mouse IgG2a (66989-1-Ig; Proteintech)
(1:750 dilution) 
Mouse IgG1 (RV202; Santa Cruz)
(1:300 dilution) 
 
Secondary ab Donkey anti-rabbit IgG (ab150075; Abcam)
(1:50 dilution) 
Rat anti-mouse IgG2a (RMG2a-62; BioLegend)
(1:20 dilution) 
Goat anti-mouse IgG1 (1071-01; Southern bio)
(1:100 dilution) 
 
Tertiary ab  Donkey anti-rat IgG (ab150153; Abcam)
(1:40 dilution) 
Donkey anti-goat IgG (ab175704; Abcam)
(1:100 dilution) 
 
Conjugate/dye AF647 AF488 AF568 DAPI 
Longpass filter 610LP 515LP 590LP 420LP 

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