Table 4.
Overview of antibodies used for on‐chip staining
| Staining with unconjugated antibodies | ||||||
|---|---|---|---|---|---|---|
| Primary antibody | Coupled with secondary antibody | |||||
| Staining target | Antibody | Specifics | Final concentration/dilution | Antibody | Specifics | Final concentration/dilution |
| Perilipin A | Anti‐Perilipin A antibody produced in rabbit | P1998 [Merck KGaA] | 12 µg mL−1 | F(ab′)2‐Goat anti‐Rabbit IgG (H+L) Cross‐Adsorbed Secondary Antibody, Alexa Fluor 555 | A‐21430 (Thermo Fisher Scientific Inc.) | 20 µg mL−1 |
| CD31 | Monoclonal Mouse Anti‐Human CD31, Endothelial Cell (Dako Omnis) | M0823 (Agilent Technologies, Inc.) | 1:50 | Goat anti‐Mouse IgG (H+L) Cross‐Adsorbed Secondary Antibody, Alexa Fluor 546 | A‐11003 (Thermo Fisher Scientific Inc.) | 20 µg mL−1 |
| CD68 | Purified Mouse Anti‐Human CD68 |
556 059 Clone Y1/82A (RUO) [BD] |
5 µg mL−1 | Goat anti‐Mouse IgG (H+L) Cross‐Adsorbed Secondary Antibody, Alexa Fluor 488 | A‐11001 (Thermo Fisher Scientific Inc.) | 20 µg mL−1 |
| CD86 | Recombinant Anti‐CD86 antibody [EPR21962] | ab239075 [Abcam] | 11.38 µg mL−1 | F(ab′)2‐Goat anti‐Rabbit IgG (H+L) Cross‐Adsorbed Secondary Antibody, Alexa Fluor 555 | A‐21430 (Thermo Fisher Scientific Inc.) | 20 µg mL−1 |
| CD206 | Recombinant Anti‐Mannose Receptor antibody [EPR6828(B)] | ab125028 [Abcam] | 3.6 µg mL−1 | F(ab′)2‐Goat anti‐Rabbit IgG (H+L) Cross‐Adsorbed Secondary Antibody, Alexa Fluor 555 | A‐21430 (Thermo Fisher Scientific Inc.) | 20 µg mL−1 |
| Trem2 | TREM2 Recombinant Rabbit Monoclonal Antibody (9H4L26) | 702 886 (Thermo Fisher Scientific Inc.) | 5 µg mL−1 | F(ab′)2‐Goat anti‐Rabbit IgG (H+L) Cross‐Adsorbed Secondary Antibody, Alexa Fluor 555 | A‐21430 (Thermo Fisher Scientific Inc.) | 20 µg mL−1 |
| Staining with conjugated antibodies | |||||
|---|---|---|---|---|---|
| Antibody | Specifics | Final concentration/dilution | Staining prior to fixation | Staining post fixation | |
| CD31 | CD31 Antibody, anti‐human, REAfinity | 130‐110‐807 (Miltenyi Biotec B.V. & Co. KG) | 1:25 | Yes | No |
| CD106 | CD106 (VCAM‐1) Antibody, anti‐human, REAfinity | 130‐104‐164 (Miltenyi Biotec B.V. & Co. KG) | 1:10 | Yes | No |
| CD309 | CD309 (VEGFR‐2) Antibody, anti‐human, REAfinity | 130‐117‐984 (Miltenyi Biotec B.V. & Co. KG) | 1:35 | Yes | No |
| eNOS | eNOS Antibody, anti‐human, APC, REAfinity | 130‐106‐840 (Miltenyi Biotec B.V. & Co. KG) | 1:10 | No | Yes |
| CD45 | CD45 Antibody, anti‐human, APC, REAfinity | 130‐110‐771 (Miltenyi Biotec B.V. & Co. KG) | 1:25–1:10 | Yes | No |
| CD11c | CD11c Antibody, anti‐human, APC, REAfinity | 130‐114‐110 (Miltenyi Biotec B.V. & Co. KG) | 1:25 | No | Yes |