ABflo® 488 Rabbit anti-Human/Monkey CD86 mAb
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Flow cytometry:1X10^6 Human PBMC were surface-stained with ABflo® 450 Rabbit anti-Human/Monkey CD14 mAb (A25196,5 μl/Test) and ABflo® 488 Rabbit IgG isotype control (A22069,5 μl/Test,left) or ABflo® 488 Rabbit anti-Human CD86 mAb (A21940,5 μl/Test,right). Cells in the monocytes gate were used for analysis.
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Flow cytometry:1X10^6 Cynomolgus PBMC were surface-stained with ABflo® 450 Rabbit anti-Human/Monkey CD14 mAb (A25196,5 μl/Test) and ABflo® 488 Rabbit IgG isotype control (A22069,5 μl/Test,left) or ABflo® 488 Rabbit anti-Human/Monkey CD86 mAb (A21940,5 μl/Test,right). Cells in the monocytes gate were used for analysis.
ABflo® 488 Rabbit anti-Human PD-L1/CD274 mAb
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Flow cytometry: 1X10^6 NCI-H460 cells treated with IFN-γ (RP01038,100 ng/mL) were surface-stained with ABflo® 488 Rabbit IgG isotype control (A22069,5 μl/Test,left) or ABflo® 488 Rabbit anti-Human PD-L1/CD274 mAb (A22304,5 μl/Test,right)
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Flow cytometry: 1X10^6 Human PBMC (untreated,left) and Human PBMC (treated with 5 µg/mL PHA for 72 hours,right) were surface-stained with APC Rabbit anti-Human/Monkey CD3 mAb (A27111,5 μl/Test) and ABflo® 488 Rabbit anti-Human PD-L1/CD274 mAb (A22304,5 μl/Test). Cells in the lymphocyte gate were used for analysis.
ABflo® 488 Rabbit anti-Human CD27 mAb
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Flow cytometry:1X10^6 Human PBMC were surface-stained with ABflo® 450 Rabbit anti-Human/Monkey CD19 mAb (A27286,5 μl/Test) and ABflo® 488 Rabbit IgG isotype control (A22069,5 μl/Test,left) or ABflo® 488 Rabbit anti-Human CD27 mAb (A22063,5 μl/Test,right). Cells in the lymphocyte gate were used for analysis.
ABflo® 488 Rabbit anti-Human CD28 mAb
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Flow cytometry:1X10^6 K-562 cells(negative control,left) and U266 cells (right) were surface-stained with ABflo® 488 Rabbit anti-Human CD28 mAb(A23405,5 μl/Test,orange line) or ABflo® 488 Rabbit IgG isotype control (A22069,5 μl/Test,blue line).Non-fluorescently stained cells was used as blank control (red line).
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Flow cytometry:1X10^6 U266 cells were surface-stained with ABflo® 488 Rabbit IgG isotype control (A22069,5 μl/Test,left) or ABflo® 488 Rabbit anti-Human CD28 mAb(A23405,5 μl/Test,right).
PE Rabbit anti-Human CD152/CTLA-4 mAb
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Flow cytometry: 1X10^6 Human PBMC (untreated,left) and Human PBMC (treated with 50 ng/mL PMA and 1 µg/mL ionomycin for 6 hours,right) were surface-stained with ABflo® 647 Rabbit anti-Human/Monkey CD4 mAb (A25279,5 μl/Test) , then intracellularly-stained with PE Rabbit anti-Human CD152/CTLA-4 mAb (A23713,5 μl/Test).
ABflo® 594 Rabbit anti-Human CD47 mAb
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Flow cytometry: 1X10^6 Human PBMC were surface-stained with ABflo® 594 Rabbit anti-Human CD47 mAb (A24302,5 μl/Test,orange line) or ABflo® 594 Rabbit IgG isotype control (A23821,5 μl/Test,blue line). Non-fluorescently stained cells were used as blank control (red line). Cells in the lymphocyte gate were used for analysis.
ABflo® 594 Rabbit anti-Mouse CD47 mAb
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Flow cytometry: 1X10^6 C57BL/6 mouse splenocytes were surface-stained with ABflo® 594 Rabbit IgG isotype control (A23821,5 μl/Test,left) or ABflo® 594 Rabbit anti-Mouse CD47 mAb (A24379,5 μl/Test,right).
Western blot analysis of lysates from wild type (WT) and PD-L1/CD274 knockout (KO) A-549 cells using PD-L1/CD274 Rabbit mAb (A19135) at 1:1000 dilution incubated overnight at 4℃. Wild type (WT) and PD-L1/CD274 knockout (KO) A549 cells were treated with IFNG (100 ng/mL) at 37℃ for 48 hours.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 30 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 45s.
